WorldWideScience

Sample records for psyllidae diaphorina citri

  1. Presencia de Diaphorina citri (Hemiptera: Psyllidae en el Noroeste Argentino (NOA Record of Diaphorina citri (Hemiptera: Psyllidae in North West Argentina

    Directory of Open Access Journals (Sweden)

    Lucrecia Augier

    2006-12-01

    Full Text Available Se amplía la distribución de Diaphorina citri Kuwayama (Hemiptera: Psyllidae para el Noroeste Argentino.The distribution of Diaphorina citri Kuwayama (Hemiptera: Psyllidae is expanded to North West Argentina.

  2. The complete mitochondrial genome sequence of Diaphorina citri (Hemiptera: Psyllidae)

    Science.gov (United States)

    The first complete mitochondrial genome (mitogenome) sequence of Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae), from Guangzhou, China is presented. The circular mitogenome is 14,996 bp in length with an A+T content of 74.5%, and contains 13 protein-coding genes (PCGs), 22 tRNA genes ...

  3. Vibrational communication between the sexes in Diaphorina citri (Hemiptera: Psyllidae)

    Science.gov (United States)

    We examined the substrate-borne vibrational signals used in communication between the sexes in Diaphorina citri Kuwayama (Hemiptera: Psyllidae), a vector of huanglongbing (an economically devastating disease of citrus), in an anechoic chamber and an olfactometer. Males and females both primarily pro...

  4. Brachygastra mellifica (Hymenoptera: Vespidae): Predation preference and feeding behavior on Diaphorina citri (Hemiptera: Psyllidae) in Mexico

    OpenAIRE

    Reyes-Rosas, M. A.; Loera-Gallardo, J.; López-Arroyo, J. I.; Buck, M.

    2014-01-01

    In previous field studies in Northern Mexico, we found the wasp Brachygastra mellifica (Say 1837) (Hymenoptera: Vespidae) preying voraciously the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), the vector of the bacteria Ca. Liberibacter spp., the putative agent of Huanglongbing, one of the most devastating citrus disease in the world.   As in Mexico, the ACP management considers the use of pest biological control, the availability of potential agents for the con...

  5. First record of Diaphorina citri (Hemiptera: Psyllidae) in Ecuador infesting urban citrus and orange jasmine trees.

    Science.gov (United States)

    Cornejo, J F; Chica, E J

    2014-01-01

    Adults and nymphs of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), were collected in the Guayaquil, Samborondón, and Durán cantons in coastal Ecuador. Psyllids were found in high numbers in citrus (Citrus spp., Sapindales: Rutaceae) and orange jasmine (Murraya exotica [L.] Jack, Sapindales: Rutaceae) trees within the Guayaquil-Samborondon-Duran conurbation; however, none was found during scoutings in the main citrus producing areas in coastal Ecuador. To the best of our knowledge, this is the first report of D. citri in Ecuador and the Pacific coastal plain of South America.

  6. Biology of Diaphorina citri (Homoptera: Psyllidae) on four host plants.

    Science.gov (United States)

    Tsai, J H; Liu, Y H

    2000-12-01

    The biology of the citrus psyllid Diaphorina citri Kuwayama was studied at 25 degrees C on four commonly grown citrus and related plants [rough lemon, Citrus jambhiri Lush; sour orange, C aurantium L.; grapefruit, C. paradisi Macfadyen; and orange jessamine, Murraya paniculata (L.) Jack] in the laboratory. The biological characteristics of each life stage are described. The average egg incubation periods on orange jessamine, grapefruit, rough lemon, and sour orange varied very little (4.1-4.2 d). The average nymphal developmental periods on these four host plants were essentially the same except the fifth stadium. Survival of immatures on orange jessamine, grapefruit, rough lemon, and sour orange was 75.4, 84.6, 78.3, and 68.6%, respectively. Female adults lived an average of 39.7, 39.7, 47.6, and 43.7 d on these respective host plants. The average number of eggs laid per female on grapefruit (858 eggs) was significantly more than those on other hosts (P < 0.05). The intrinsic rate of natural increase (r(m)) for D. citri on grapefruit was highest. Jackknife estimates of r(m) varied from 0.188 on grapefruit to 0.162 on orange jessamine and rough lemon. The mean population generation time on these hosts ranged from 31.6 to 34.1 d. The continuous flushes produced by orange jessamine could play an important role in maintaining high populations of this vector when the new flushes are not available in the commercial citrus groves.

  7. Population ecology and phenology of Diaphorina citri (Hemiptera: Psyllidae) in two Florida citrus groves

    Science.gov (United States)

    Studies were conducted during 2005 and 2006 to assess population densities and phenology of the psyllid Diaphorina citri Kuwayama in citrus within east central Florida. Young, irrigated grapefruit and mature, non-irrigated orange trees were sampled weekly for eggs, nymphs and adults on flush shoots;...

  8. The effects of host, geographic origin, and gender on the thermal requirements of Diaphorina citri (Hemiptera: Psyllidae).

    Science.gov (United States)

    Nava, Dori E; Gomez-Torres, Mariuxi L; Rodrigues, Marjorie D; Bento, José M S; Haddad, Marinéia L; Parra, José R P

    2010-04-01

    Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the vector of the bacteria that causes citrus greening and is considered one of the world's most important citrus diseases. We examined how host, geographic region, and gender affect the thermal requirements of D. citri. The insects were reared in climatic chambers at constant temperatures of 18, 20, 22, 25, 28, 30, and 32 +/- 1 degrees C, 70 +/- 10% RH, and a 14 h photophase. Host plants for D. citri included orange (Citrus sinensis [Rutaceae]) varieties Pêra and Natal, the rootstock, Rungpur lime (C. limonia [Rutaceae]) and the natural host, Orange jessamine (Murraya paniculata [Rutaceae]). To study the influence of geographic origin on thermal requirements, we studied D. citri populations from Piracicaba, SP (warmer region) and Itapetininga, SP (cooler region). The duration and survival of the development stages and the duration of the total development (egg-adult) did not differ significantly on the different hosts, but it did vary with temperature. Nymphs of D. citri created on the different hosts have the same thermal requirements. The thermal requirements for this species collected from the two climate regions were identical; males and females also had the same thermal requirements.

  9. Spatial distribution of Diaphorina citri Kuwayama (Hemiptera: Psyllidae in citrus orchards Distribuição espacial de Diaphorina citri Kuwayama (Hemiptera: Psyllidae em pomares de citros

    Directory of Open Access Journals (Sweden)

    Marilia Gregolin Costa

    2010-10-01

    Full Text Available The psyllid Diaphorina citri Kuwayama is one of the most important pests of citrus, mainly because it is the vector of the bacterium that causes huanglongbing (HLB or 'Greening' disease. To study the spatial distribution of nymphs and adults of this pest, an experiment was carried out in two 'Valencia' sweet orange orchards, four and 12 years of age, established in Matão, central area of São Paulo state, Brazil. The following dispersion indices were used to study pest aggregation in the citrus plants: variance/mean relationship (I, index of Morisita (Iδ, coefficient of Green (Cx, the exponent of negative binomial distribution, common k (c and Taylor's Power Law for each sampling. The negative binomial distribution was more representative of the spatial distribution of this psyllid, for both nymphs and adults. For most samplings, psyllid nymphs found in branches and adults caught in traps had an aggregated distribution.O psilídeo Diaphorina citri Kuwayama tornou-se nos últimos anos uma das mais importantes pragas na cultura de citros, principalmente pelos prejuízos causados às plantas por ser o transmissor da bactéria causadora da doença Huanglongbing (HLB ou 'Greening'. Com a finalidade de estudar a distribuição espacial de ninfas e adultos desta praga, instalaram-se experimentos em duas áreas de citros com histórico de ocorrência de HLB, no município de Matão (região central do Estado de São Paulo, em plantas de laranja 'Valência', com quatro e 12 anos de idade. Para estudo da agregação da população nas plantas, foram utilizados os seguintes índices de dispersão: razão variância/média (I, índice de Morisita (Iδ, coeficiente de Green (Cx e expoente da distribuição binomial negativa, k comum (c e lei da potência de Taylor para cada amostragem. A distribuição binomial negativa foi o modelo mais adequado para representar a distribuição espacial do psilídeo, tanto para ninfas como para adultos. Na maioria das

  10. Pengaruh Jenis Mangsa dan Suhu pada Perkembangan Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae dan Peranannya dalam Pengendalian Diaphorina citri Kuwayama (Hemiptera: Psyllidae

    Directory of Open Access Journals (Sweden)

    Tris Haris Ramadhan

    2008-07-01

    Full Text Available Diaphorina citri Kuwayama (Hemiptera: Psyllidae is the vector of citrus greening (Huanglongbing bacterium and the most serious impediment to citrus culture. Classical biological control of this psyllid vector should contribute to suppress their population. This research was conducted to determine the performance of Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae when they were fed with D. citri. The larval performance index of M. sexmaculatus on D. citri compared with Aphis craccivora Koch (Hemiptera: Aphididae diet was 1.3.M. sexmaculatus fed with D. citri had lower fitness than those fed with A. craccivora as shown by longer larval stadium, lower adult dry weight, less number of egg produced and lower percentage of egg hatched. M. sexmaculatus grew best at the temperature of 27oC. Employing the exclusion procedure under field condition,M. sexmaculatus could reduce the population of D. citri up to 90%. These findings showed that theM. sexmaculatus could be a potential predator in reducing D. citri, particularly when the more preferred prey A. craccivora was not present.   Diaphorina citri Kuwayama (Hemiptera: Psyllidae merupakan kelompok Psyllid yang menularkan penyebab penyakit Huanglongbing yang sangat berbahaya pada tanaman jeruk. Pengendalian hayati klasik telah banyak memberikan kontribusi dalam pengendalian di lapangan. Penelitian ini dilakukan untuk melihat penampilan Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae dengan pakan D. citri. Indeks penampilan larva M. sexmaculatus dengan pakan D. citri dibandingkan dengan Aphis craccivora Koch (Hemiptera: Aphididae adalah 1,3. M. sexmaculatus yang diberi pakan D. citri menunjukkan penurunan kebugaran dibandingkan jika diberi pakan A. craccivora seperti yang ditunjukkan dengan stadium larva lebih lama, penurunan berat kering serangga dewasa, telur yang dihasilkan lebih sedikit, dan penurunan jumlah telur yang menetas. Menggunakan metode eksklusi pada kondisi

  11. Evaluation of six different groups of insecticides for the control of citrus psylla Diaphorina citri (Hemiptera: Psyllidae

    Directory of Open Access Journals (Sweden)

    Rakhmin Gul

    2005-01-01

    Full Text Available Studies on the efficacy of different insecticides against citrus psylla, Diaphorina citri (Hemiptera: Psyllidae were carried out at Agricultural Research Institute, Tarnab, Peshawar, Pakistan. Six insecticides viz. Actara 25 WG, (thiamethoxam Cascade 10 DC (Flufenoxuron, Match 050 EC (lufenuron, Thiodan 35 EC (endosulfan, Karate 2.5 EC (α-cyhalothrin, and Supracide 40 EC (methidathion, were tested for their effectiveness against D. citri. After first spray overall mean population of D. citri was 3.63, 4.75, 5.59, 6.66, 7.47, 8.11 per six inches tender shoot on Actara 25 WG, Cascade 10 DC, Match 050 EC, Thiodan 35 EC, Karate 2.5 EC and Supracide 40 EC treated plants respectively, while on control plants the population was 12.39. Similarly, after the second spray of each of the same insecticides the population of D. citri was 2.65, 4.23, 5.61, 6.41, 7.35 and 8.73 respectively. Where in controls there were 15.18 psyllids. Percent decrease of D. citri population in comparison to control after the first spray was highest in Actara 25 WG (72.20 followed by Cascade 10 DC (62.91, Match 050 EC (54.07, Thiodan 35 EC (47.61, Karate 2.5 EC (38.94 and Supracide 40 EC (35.74. After the second spray percent decrease over control recorded was highest in Actara 25 WG (83.54, followed by Cascade 10 DC (71.08, Match 050 EC (63.94, Thiodan 35 EC (60.79, Karate 2.5 EC (52.52 and Supracide 40 EC (45.62.

  12. Insecticide sprays, natural enemy assemblages and predation on Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae).

    Science.gov (United States)

    Monzo, C; Qureshi, J A; Stansly, P A

    2014-10-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is considered a key citrus pest due to its role as vector of 'huanglongbing' (HLB) or citrus greening, probably the most economically damaging disease of citrus. Insecticidal control of the vector is still considered a cornerstone of HLB management to prevent infection and to reduce reinoculation of infected trees. The severity of HLB has driven implementation of intensive insecticide programs against ACP with unknown side effects on beneficial arthropod fauna in citrus agroecosystems. We evaluated effects of calendar sprays directed against this pest on natural enemy assemblages and used exclusion to estimate mortality they imposed on ACP populations in citrus groves. Predator exclusion techniques were used on nascent colonies of D. citri in replicated large untreated and sprayed plots of citrus during the four major flushing periods over 2 years. Population of spiders, arboreal ants and ladybeetles were independently assessed. Monthly sprays of recommended insecticides for control of ACP, adversely affected natural enemy populations resulting in reduced predation on ACP immature stages, especially during the critical late winter/early spring flush. Consequently, projected growth rates of the ACP population were greatest where natural enemies had been adversely affected by insecticides. Whereas, this result does not obviate the need for insecticidal control of ACP, it does indicate that even a selective regimen of sprays can impose as yet undetermined costs in terms of reduced biological control of this and probably other citrus pests.

  13. Entomophagous insects associated to Diaphorina citri (Hemiptera: Psyllidae) in citrus orchards with different weed management systems in Papantla, Veracruz, Mexico

    OpenAIRE

    Ortega-Arenas, L. D.; López-López, R.; Lomelí-Flores, J. R.; Cedillo-Portugal, E.; Gómez-Tovar, L.; Salazar-Cruz, J.; Villegas-Monter, A.

    2014-01-01

    Huanglongbing (HLB), one of the most destructive diseases of citrus worldwide, is threatening the survival of the citrus industry in Mexico. Diaphorina citri is the primary vector of HLB; thus, control of the vector it’s vital for disease management. This study was carried out to evaluate the influence of different management systems on the population psyllid density and entomophagous insects associated in orange orchards (Citrus sinensis cv. Valencia) in Papantla, Veracruz, Mexico. Five orch...

  14. Roles of olfactory cues, visual cues, and mating status in orientation of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) to four different host plants.

    Science.gov (United States)

    Wenninger, Erik J; Stelinski, Lukasz L; Hall, David G

    2009-02-01

    Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important worldwide pest of citrus that vectors bacteria (Candidatus Liberibacter spp.) responsible for huanglongbing (citrus greening disease). We examined the behavioral responses of mated and unmated D. citri of both sexes to odors from host plants in a Y-tube olfactometer, with and without visual cues. The host plants tested were 'Duncan' grapefruit (Citrus paradisi Macfayden), sour orange (Citrus aurantium L.), navel orange (C. sinensis L.), and Murraya paniculata L. Jack. Responses varied by plant species, psyllid sex and mating status, and the presence of a visual cue. Evidence of attraction generally was stronger in females and in mated individuals of both sexes relative to virgins. The presence of a visual cue typically enhanced attractiveness of olfactory cues; in no case did unmated individuals show evidence of attraction to host plant odors in the absence of avisual cue. In the absence of visual cues, mated females and males showed evidence of attraction only to odors from sour orange and navel orange, respectively. Psyllids exhibited anemotactic responses when assayed with plant odors alone but showed strong evidence of attraction only when olfactory and visual cues were combined, suggesting that olfactory cues facilitate orientation to host plants but may be insufficient alone. Antennal responses to citrus volatiles were confirmed by electroantennogram. The results reported here provide evidence that D. citri uses olfactory and visual cues in orientation to host plants and suggest the possibility of using plant volatiles in monitoring and management of this pest.

  15. Behavioral, ultrastructural and chemical studies on the honeydew and waxy secretions by nymphs and adults of the Asian citrus psyllid Diaphorina citri (Hemiptera: Psyllidae.

    Directory of Open Access Journals (Sweden)

    El-Desouky Ammar

    Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri (Hemiptera: Psyllidae is the primary vector of the bacterium causing citrus huanglongbing (citrus greening, the most serious disease of citrus worldwide. Psyllids and other hemipterans produce large amounts of honeydew, which has been used previously as an indicator of phloem sap composition and insect feeding or metabolism. Behavioral, ultrastructural and chemical studies on ACP, its honeydew and waxy secretions showed important differences between nymphs, males and females, and suggested some mechanisms by which the psyllids, especially nymphs and adult females, can minimize their contamination with honeydew excretions. The anal opening in ACP, near the posterior end of the abdomen, is on the ventral side in nymphs and on the dorsal side in adult males and females. Video recordings showed that adult males produce clear sticky droplets of honeydew gently deposited behind their body on the leaf surface, whereas adult females produce whitish honeydew pellets powerfully propelled away from the female body, probably to get their excretions away from eggs and newly hatched nymphs. ACP nymphs produce long ribbons or tubes of honeydew that frequently stay attached to the exuviae after molting, or drop when feeding on the lower side of citrus leaves. Furthermore, honeydew excretions of both nymphs and adult females are covered with a thin layer of whitish waxy material ultrastructurally composed of a convoluted network of long fine filaments or ribbons. This material is extruded from intricate arrays of wax pores in the circumanal ring (around the anus that is found in nymphs and females but not in males of ACP or other psyllid species. Infrared microscopy and mass spectroscopy revealed that, in addition to various sugars, honeydew excretions of ACP nymphs and females are covered with a thin layer of wax similar in profile to ester waxes.

  16. Control químico de Diaphorina citri Kuwayama (Hemíptera: Psyllidae en lima persa Citrus latifolia Tanaka Chemical control of Diaphorina citri Kuwayama (Hemiptera: Psyllidae in Persian lime Citrus latifolia Tanaka

    Directory of Open Access Journals (Sweden)

    Luis Martín Hernández-Fuentes

    2012-06-01

    Full Text Available A nivel mundial, la citricultura representa una actividad de gran importancia. México ocupa el cuarto lugar mundial en producción. Debido a la introducción y dispersión del vector D. citri en todas las zonas citrí colas de México y la reciente detección de la enfermedad conocida como huanglongbing causada por la bacteria Candidatus Liberibacter asiaticus, es necesaria la evaluación de métodos de control de este insecto. En el año 2010 en Nayarit, México, se realizaron pruebas de efectividad técnica en campo y persistencia de productos químicos de diferente mecanismo de acción contra D. citri en lima persa. Se hicieron dos evaluaciones en fechas distintas. En cada ensayo se evaluó número de ninfas por brote y porcentaje de brotes infestados. De los productos evaluados el dimetoato, imidacloprid y la mezcla de imidacloprid más betaciflutrina ejercieron más de 85 % de control de ninfas durante 27 días después de la aplicación. En los árboles tratados con dimetoato e imidacloprid en dosis de 400 mL y 300 mL ha-¹, respectivamente, se observó 100% de brotes libres de ninfas de D. citri a los 27 días después de la aplicación. El aceite agrícola en dosis de dos y tres L ha-1 ejerció menor control de ninfas de D. citri.Citrus crops represent an outstanding activity at worldwide level. Mexico ranks fourth place in production. Due introduction and dispersion of D. citri vector in all citric zones from Mexico and the newly detected disease known as huanglongbing (HLB caused by Candidatus Liberibacter asiaticus bacteria, assessment of control methods for this bug is required. In 2010, technical effectiveness in field and persistence for different action route against Persian lime D. citri chemical products tests were performed in Nayarit, Mexico. Two assessments were made at different dates. The amount of nymphs per shoot and percentage of infested shoots were assessed in each rehearsal. From evaluated products, dimethoate

  17. Control químico de Diaphorina citri Kuwayama (Hemíptera: Psyllidae) en lima persa Citrus latifolia Tanaka Chemical control of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) in Persian lime Citrus latifolia Tanaka

    OpenAIRE

    Luis Martín Hernández-Fuentes; Mario Alfonso Urias-López; José Isabel López-Arroyo; Rafael Gómez-Jaimes; Néstor Bautista-Martínez

    2012-01-01

    A nivel mundial, la citricultura representa una actividad de gran importancia. México ocupa el cuarto lugar mundial en producción. Debido a la introducción y dispersión del vector D. citri en todas las zonas citrí colas de México y la reciente detección de la enfermedad conocida como huanglongbing causada por la bacteria Candidatus Liberibacter asiaticus, es necesaria la evaluación de métodos de control de este insecto. En el año 2010 en Nayarit, México, se realizaron pruebas de efectividad t...

  18. Natural parasitism of Diaphorina citri Kuwayama (Hemiptera, Psyllidae nymphs by Tamarixia radiata Waterston (Hymenoptera, Eulophidae in São Paulo orange groves

    Directory of Open Access Journals (Sweden)

    Paulo Eduardo Branco Paiva

    2012-12-01

    Full Text Available The psyllid Diaphorina citri Kuwayama 1908 has become the main citrus pest species in the state of São Paulo, Brazil, after the introduction of the huanglongbing or citrus greening. This study evaluated the parasitism of 3rd, 4th and 5th instar D. citri nymphs by Tamarixia radiata (Waterston, 1922 in citrus groves under a regimen of regular insecticide applications in ten producing regions: Araraquara, Barretos, Bauru, Botucatu, Franca, Itapetininga, Jaú, Limeira, Lins and São João da Boa Vista. Sixty-nine samples of new branches infested with nymphs of D. citri were collected from 2005 to 2008 in orange groves ranging from 1 to 20 years old, of the varieties Hamlin, Pera, Valencia and Natal. The parasitoid T. radiata is widely distributed in São Paulo orange groves, and was identified in 50 (72% of the samples, showing a mean parasitism rate of 12.4%. The highest parasitism rate was observed in the "summer" (from January through March, with a mean of 25.7%. Nymphal parasitism was above 90% in two samples. The probable causes of the variations in parasitism of D. citri by T. radiata are discussed.

  19. Gender differences and effect of photophase on Asian citrus psyllid (Diaphorina citri Kuwayama) feeding behavior

    Science.gov (United States)

    Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), thought to be primarily a phloem-feeding insect, transmits the presumptive pathogen for Huanglongbing, ‘Candidatus Liberibacter asiaticus’. Because this bacterium is restricted to the phloem and bacterial transmission is the res...

  20. Chemical and behavioral analysis of the cuticular hydrocarbons from Asian citrus psyllid, Diaphorina citri

    Science.gov (United States)

    Huanglongbing (HLB) is the most destructive disease of citrus worldwide. Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the vector of the phloem-inhabiting bacterium, Candidatus Liberibacter asiaticus, which is presumed to cause HLB. Laboratory and field studies were cond...

  1. Geographic Distribution of Habitat, Development, and Population Growth Rates of the Asian Citrus Psyllid, Diaphorina citri, in Mexico

    OpenAIRE

    López-Collado, José; Isabel López-Arroyo, J.; Robles-García, Pedro L.; Márquez-Santos, Magdalena

    2013-01-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is an introduced pest in Mexico and a vector of huanglongbing, a lethal citrus disease. Estimations of the habitat distribution and population growth rates of D. citri are required to establish regional and areawide management strategies and can be used as a pest risk analysis tools. In this study, the habitat distribution of D. citri in Mexico was computed with MaxEnt, an inductive, machine-learning program that uses...

  2. Natural parasitism of Diaphorina citri Kuwayama (Hemiptera, Psyllidae nymphs by Tamarixia radiata Waterston (Hymenoptera, Eulophidae in São Paulo orange groves Parasitismo natural de ninfas de Diaphorina citri Kuwayama (Hemiptera, Psyllidae por Tamarixia radiata Waterston (Hymenoptera, Eulophidae em pomares de laranja em São Paulo

    Directory of Open Access Journals (Sweden)

    Paulo Eduardo Branco Paiva

    2012-12-01

    Full Text Available The psyllid Diaphorina citri Kuwayama 1908 has become the main citrus pest species in the state of São Paulo, Brazil, after the introduction of the huanglongbing or citrus greening. This study evaluated the parasitism of 3rd, 4th and 5th instar D. citri nymphs by Tamarixia radiata (Waterston, 1922 in citrus groves under a regimen of regular insecticide applications in ten producing regions: Araraquara, Barretos, Bauru, Botucatu, Franca, Itapetininga, Jaú, Limeira, Lins and São João da Boa Vista. Sixty-nine samples of new branches infested with nymphs of D. citri were collected from 2005 to 2008 in orange groves ranging from 1 to 20 years old, of the varieties Hamlin, Pera, Valencia and Natal. The parasitoid T. radiata is widely distributed in São Paulo orange groves, and was identified in 50 (72% of the samples, showing a mean parasitism rate of 12.4%. The highest parasitism rate was observed in the "summer" (from January through March, with a mean of 25.7%. Nymphal parasitism was above 90% in two samples. The probable causes of the variations in parasitism of D. citri by T. radiata are discussed.O psilídeo Diaphorina citri Kuwayama 1908 tornou-se a principal praga dos citros no estado de São Paulo após a introdução do huanglongbing ou grenning dos citros. Este estudo avaliou as proporções de ninfas de terceiro a quinto ínstares de D. citri parasitadas por Tamarixia radiata (Waterston, 1922 em pomares de laranja submetidos a pulverizações constantes de inseticidas em dez regiões produtoras, Araraquara, Barretos, Bauru, Botucatu, Franca, Itapetininga, Jaú, Limeira, Lins e São João da Boa Vista. Foram coletadas 69 amostras de ramos jovens infestados com ninfas de D. citri obtidas entre 2005 e 2008 em pomares de 1 a 20 anos de idade, das cultivares Hamilin, Pera, Valencia e Natal. O parasitóide T. radiata encontra-se amplamente distribuído em pomares de São Paulo, tendo sido observado em 50 amostras (72%, com taxa média de

  3. Sequencing and annotation of the Wolbachia endosymbiont of Diaphorina citri by the CG-HLB genome resources group reveals candidate sources of interaction with the insect host

    Science.gov (United States)

    Diaphorina citri (Hemiptera: Psyllidae), the Asian citrus psyllid, is the vector of Ca. Liberibacter asiaticus, the causal agent of huanglongbing. The D. citri metagenome has been completed to gain a better understanding of the biology of this organism and the potential roles of other psyllid bacter...

  4. Use of the entomopathogenic fungi Metarhizium anisopliae, Cordyceps bassiana and Isaria fumosorosea to control Diaphorina citri (Hemiptera: Psylidae) in Persian lime under field conditions

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is a destructive insect pest in the citriculture, because it is an efficient vector of the proteobacteria, ‘Candidatus Liberibacter asiaticus’ (Las), ‘Ca. L. Africanus’ (Laf), and ‘Ca. L. Americanus’ (Lam). These bacteria c...

  5. Temporal response and attraction of Diaphorina citri to visual stimuli

    Science.gov (United States)

    As the vector of the global disease of citrus greening or huanglongbing, Asian citrus pysllids, Diaphorina citri Kuwayama (Hemiptera; Liviidae) are the greatest threat to the worldwide citrus industry. Critical to management of D. citri and huanglongbing, is optimization of surveillance methodologie...

  6. Chemical and behavioral analysis of the cuticular hydrocarbons from Asian citrus psyllid, Diaphorina citri.

    Science.gov (United States)

    Mann, Rajinder S; Rouseff, Russell L; Smoot, Jack; Rao, Nandikeswara; Meyer, Wendy L; Lapointe, Stephen L; Robbins, Paul S; Cha, Dong; Linn, Charles E; Webster, Francis X; Tiwari, Siddharth; Stelinski, Lukasz L

    2013-06-01

    Huanglongbing (HLB) is the most destructive disease of citrus worldwide. The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the vector of the phloem-inhabiting bacterium, Candidatus Liberibacter asiaticus, which is presumed to cause HLB in Florida citrus. Laboratory and field studies were conducted to examine the behavioral responses of male and female D. citri to their cuticular extracts. In olfactometer assays, more male D. citri were attracted to one, five, or 10 female cuticular extract equivalent units than blank controls. The results were confirmed in field studies in which clear or yellow traps baited with 10 female cuticular extract equivalent units attracted proportionately more males than clear traps baited with male cuticular extract or unbaited traps. Analyses of cuticular constituents of male and female D. citri revealed differences between the sexes in chemical composition of their cuticular extracts. Laboratory bioassays with synthetic chemicals identified from cuticular extracts indicated that dodecanoic acid attracted more males than clean air. Traps baited with dodecanoic acid did not increase total catch of D. citri as compared with blank traps at the dosages tested; however, the sex ratio of psyllid catch was male biased on traps baited with the highest lure loading dosage tested (10.0 mg). © 2012 The Authors Insect Science © 2012 Institute of Zoology, Chinese Academy of Sciences.

  7. Construction and evaluation of yeast two-hybrid cDNA library of Diaphorina citri (Hemiptera: Psyllidae)%柑橘木虱酵母双杂交cDNA文库的构建及评价

    Institute of Scientific and Technical Information of China (English)

    马晓芳; 陈国庆; 张学潮; 徐海君

    2013-01-01

    In order to study the interacting proteins between the Asian citrus psyllid ( Diaphorina citri) and Candidatus Liberibacter asiaticus ( CLas) which is the pathogenic bacterium causing Huanglongbing, yeast two-hybrid cDNA library of D. citri was constructed using the Switching Mechanism at 5' End of the RNA Transcript (SMART) technique. The total RNA was isolated from the citrus psyllid adults bred in the laboratory and subjected to reverse transcription, and the double-strand cDNAs ( ds cDNAs) were synthesized. The ds cDNAs were ligated with homologous adapter and purified by the chromatography column. By using homologous recombination reaction, the ds cDNAs were transformed into the Y187 competent cell with the library plasmid pGADT7-Rec to construct yeast two-hybrid cDNA library. Detection of the library indicated that it contained more than 106 independent clones, the titer of the amplified library was 2. 23 × 10 cfu/mL, and the average size of inserts was above 750 bp in the cDNA library. These results demonstrate that the library meets the requirements of the standard cDNA library. Moreover, two membrane proteins, ORF420 and ORF3420, from ( CLas) were used as bait proteins to screen the interacting proteins in the library, but no positive clone was screened in the tests. The yeast two-hybrid cDNA library of D. citri will be useful for the research on the interaction between insect vectors and C. Liberibacter asiaticus in the future.%为了探索研究柑橘木虱Diaphorina citri与柑橘黄龙病(Huanglongbing,HLB)病原菌的相互作用蛋白,本研究运用RNA转录中5 '末端转换机制(Switching Mechanism at 5'End of the RNA Transcript,SMART)技术构建了柑橘木虱的酵母双杂交cDNA文库.以实验室饲养的柑橘木虱为材料,提取总RNA,经反转录后合成ds cDNA,两端添加同源重组序列,并用层析柱纯化;ds cDNA与文库质粒pGADT7-Rec在酵母Y187感受态细胞内发生同源重组,柑橘木虱cDNA重组到文库质粒

  8. Chemical and behavioral analysis of the cuticular hydrocarbons from Asian citrus psyllid, Diaphorina citri

    Institute of Scientific and Technical Information of China (English)

    Raiinder S.Mann; Russell L.Rouseff; Jack Smoot; Nandikeswara Rao; Wendy L.Meyer; Stephen L.Lapointe; Paul S.Robbins

    2013-01-01

    Huanglongbing (HLB) is the most destructive disease of citrus worldwide.The Asian citrus psyllid,Diaphorina citri Kuwayama (Hemiptera:Psyllidae),is the vector of the phloem-inhabiting bacterium,Candidatus Liberibacter asiaticus,which is presumed to cause HLB in Florida citrus.Laboratory and field studies were conducted to examine the behavioral responses of male and female D.citri to their cuticular extracts.In olfactometer assays,more male D.citri were attracted to one,five,or 10 female cuticular extract equivalent units than blank controls.The results were confirmed in field studies in which clear or yellow traps baited with 10 female cuticular extract equivalent units attracted proportionately more males than clear traps baited with male cuticular extract or unbaited traps.Analyses of cuticular constituents of male and female D.citri revealed differences between the sexes in chemical composition of their cuticular extracts.Laboratory bioassays with synthetic chemicals identified from cuticular extracts indicated that dodecanoic acid attracted more males than clean air.Traps baited with dodecanoic acid did not increase total catch of D.cirri as compared with blank traps at the dosages tested; however,the sex ratio of psyllid catch was male biased on traps baited with the highest lure loading dosage tested (10.0 mg).

  9. Complete Genome Sequence of the Largest Known Flavi-Like Virus, Diaphorina citri flavi-like virus, a Novel Virus of the Asian Citrus Psyllid, Diaphorina citri.

    Science.gov (United States)

    Matsumura, Emilyn E; Nerva, Luca; Nigg, Jared C; Falk, Bryce W; Nouri, Shahideh

    2016-09-08

    A novel flavi-like virus tentatively named Diaphorina citri flavi-like virus (DcFLV) was identified in field populations of Diaphorina citri through small RNA and transcriptome sequencing followed by reverse transcription (RT)-PCR. We report here the complete nucleotide sequence and genome organization of DcFLV, the largest flavi-like virus identified to date.

  10. Complete Genome Sequence of the Largest Known Flavi-Like Virus, Diaphorina citri flavi-like virus, a Novel Virus of the Asian Citrus Psyllid, Diaphorina citri

    Science.gov (United States)

    Matsumura, Emilyn E.; Nerva, Luca; Nigg, Jared C.; Falk, Bryce W.

    2016-01-01

    A novel flavi-like virus tentatively named Diaphorina citri flavi-like virus (DcFLV) was identified in field populations of Diaphorina citri through small RNA and transcriptome sequencing followed by reverse transcription (RT)-PCR. We report here the complete nucleotide sequence and genome organization of DcFLV, the largest flavi-like virus identified to date. PMID:27609921

  11. Survey of endosymbionts in the Diaphorina citri metagenome and assembly of a Wolbachia wDi draft genome.

    Directory of Open Access Journals (Sweden)

    Surya Saha

    Full Text Available Diaphorina citri (Hemiptera: Psyllidae, the Asian citrus psyllid, is the insect vector of Ca. Liberibacter asiaticus, the causal agent of citrus greening disease. Sequencing of the D. citri metagenome has been initiated to gain better understanding of the biology of this organism and the potential roles of its bacterial endosymbionts. To corroborate candidate endosymbionts previously identified by rDNA amplification, raw reads from the D. citri metagenome sequence were mapped to reference genome sequences. Results of the read mapping provided the most support for Wolbachia and an enteric bacterium most similar to Salmonella. Wolbachia-derived reads were extracted using the complete genome sequences for four Wolbachia strains. Reads were assembled into a draft genome sequence, and the annotation assessed for the presence of features potentially involved in host interaction. Genome alignment with the complete sequences reveals membership of Wolbachia wDi in supergroup B, further supported by phylogenetic analysis of FtsZ. FtsZ and Wsp phylogenies additionally indicate that the Wolbachia strain in the Florida D. citri isolate falls into a sub-clade of supergroup B, distinct from Wolbachia present in Chinese D. citri isolates, supporting the hypothesis that the D. citri introduced into Florida did not originate from China.

  12. Complete Genome Sequence of a Putative Densovirus of the Asian Citrus Psyllid, Diaphorina citri.

    Science.gov (United States)

    Nigg, Jared C; Nouri, Shahideh; Falk, Bryce W

    2016-07-28

    Here, we report the complete genome sequence of a putative densovirus of the Asian citrus psyllid, Diaphorina citri Diaphorina citri densovirus (DcDNV) was originally identified through metagenomics, and here, we obtained the complete nucleotide sequence using PCR-based approaches. Phylogenetic analysis places DcDNV between viruses of the Ambidensovirus and Iteradensovirus genera.

  13. Geographic distribution of habitat, development, and population growth rates of the Asian citrus psyllid, Diaphorina citri, in Mexico.

    Science.gov (United States)

    López-Collado, José; Isabel López-Arroyo, J; Robles-García, Pedro L; Márquez-Santos, Magdalena

    2013-01-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is an introduced pest in Mexico and a vector of huanglongbing, a lethal citrus disease. Estimations of the habitat distribution and population growth rates of D. citri are required to establish regional and areawide management strategies and can be used as a pest risk analysis tools. In this study, the habitat distribution of D. citri in Mexico was computed with MaxEnt, an inductive, machine-learning program that uses bioclimatic layers and point location data. Geographic distributions of development and population growth rates were determined by fitting a temperature-dependent, nonlinear model and projecting the rates over the target area, using the annual mean temperature as the predictor variable. The results showed that the most suitable regions for habitat of D. citri comprise the Gulf of Mexico states, Yucatán Peninsula, and areas scattered throughout the Pacific coastal states. Less suitable areas occurred in northern and central states. The most important predictor variables were related to temperature. Development and growth rates had a distribution wider than habitat, reaching some of the northern states of México. Habitat, development, and population growth rates were correlated to each other and with the citrus producing area. These relationships indicated that citrus producing states are within the most suitable regions for the occurrence, development, and population growth of D. citri, therefore increasing the risk of huanglongbing dispersion.

  14. Complete Genome Sequence of Diaphorina citri-associated C virus, a Novel Putative RNA Virus of the Asian Citrus Psyllid, Diaphorina citri.

    Science.gov (United States)

    Nouri, Shahideh; Salem, Nidà; Falk, Bryce W

    2016-07-21

    We present here the complete nucleotide sequence and genome organization of a novel putative RNA virus identified in field populations of the Asian citrus psyllid, Diaphorina citri, through sequencing of the transcriptome followed by reverse transcription-PCR (RT-PCR). We tentatively named this virus Diaphorina citri-associated C virus (DcACV). DcACV is an unclassified positive-sense RNA virus.

  15. GENETIC Diaphorina citri DIVERSITY ON CITRUS CROPS OF THE VALLE DEL CAUCA AND QUINDÍO (COLOMBIA

    Directory of Open Access Journals (Sweden)

    MIGUEL ANGEL MONCAYO-DONOSO

    2014-07-01

    Full Text Available The Asiatic psyllid Diaphorina citri (Hemiptera: Psyllidae is the main vector of Candidatus liberibacter, which causes the Huanglongbing HLB disease, known for devastating citrus in the world but not yet reported in Colombia. The genetic variability of the D. citri population was studied through sequencing the COI mitochondrial gene as molecular marker. Adults were collected in citrus producing zones of the Colombian Valle del Cauca and Quindío. Amplification was performed with two pairs of specific primers for Hemiptera. The PCR products were sequenced at Macrogen-Korea, obtaining a total of 124 sequences. For the bioinformatic analysis, the Vector NTI 11.5, Harlequin V 3.5, MEGA 5 and MAFFT 6 programs were used. The molecular diversity indices between populations were similar, revealing a common origin and a recent split of the populations excluding a significant genetic differentiation associated to variations of the bacterium, however the haplotype diversity index was higher than the nucleotide diversity index. The latter one showed a low number of polymorphic sites, indicating that the D. citri populations are expanding. The study of the vector’s genetic variability is a tool for the prediction of likely scenarios for the spread of diseases.

  16. Daily and seasonal patterns in abdomen color in Diaphoria citri (Hemiptera: Psyllidae)

    Science.gov (United States)

    Diaphorina citri Kuwayama, a psyllid vector of huanglongbing (citrus greening disease), exhibits three more or less distinct abdomen colors in the adult psyllid: gray/brown, blue/green, and orange/yellow. We explored the daily (in individuals in the laboratory) and seasonal (in a field population) p...

  17. Survey of endosymbionts in the Diaphorina citri metegenome and assembly of a Wolbachia wDi draft genome

    Science.gov (United States)

    Diaphorina citri, the Asian citrus psyllid,(Hemiptera) is the insect vector of Candidatus Liberibacter asiaticus, the presumed cause of citrus greening disease, known as huanglongbing. Sequencing of the Diaphorina citri metagenome has been initiated to gain better understanding of the biology of D. ...

  18. Patogenicidade de Beauveria bassiana ao psilídeo Diaphorina citri e compatibilidade do fungo com produtos fitossanitários Beauveria bassiana pathogenicity to Diaphorina citri and compatibility of the fungus with phytosanitary products

    Directory of Open Access Journals (Sweden)

    Ana Paula Ferreira Pinto

    2012-12-01

    Full Text Available O objetivo deste trabalho foi avaliar a patogenicidade de Beauveria bassiana a ninfas de Diaphorina citri (Hemiptera: Psyllidae e verificar a compatibilidade do fungo com produtos fitossanitários e sua persistência em plantas de citros. Ninfas de D. citri foram pulverizadas com B. bassiana, nas concentrações 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 e 1x10(9 conídios mL-1, para determinação da concentração letal. Para avaliação da compatibilidade do fungo com produtos fitossanitários, extrato de nim e cinco inseticidas de quatro grupos químicos diferentes foram incorporados individualmente ao meio de cultura BDA em que o fungo foi cultivado. Avaliaram-se o crescimento vegetativo, a esporulação e a viabilidade do entomopatógeno. Plantas de citros, mantidas em casa de vegetação, foram tratadas primeiramente com os produtos fitossanitários e depois com o entomopatógeno. Avaliaram-se os tempos de exposição de 24 horas e de 7 e 14 dias. O fungo foi patogênico às ninfas de D. citri; a CL50 foi de 0,4x10(7 e a CL90 de 6,7x10(7 conidios mL-1, no décimo dia de avaliação. Em laboratório, os produtos fitosssanitários reduzem o crescimento do fungo. Em casa de vegetação, os produtos não afetam a sobrevivência do fungo nas plantas de citros.The objective of this work was to evaluate the pathogenicity of Beauveria bassiana to Diaphorina citri (Hemiptera: Psyllidae nymphs, and to check the compatibility of the fungus with phytosanitary products, and its persistence in citrus plants. Nymphs of D. citri were sprayed with B. bassiana in the concentrations 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 and 1x10(9 conidia mL-1 for lethal concentration determination. In order to evaluate the compatibility of the phytosanitary products with the fungus, neem extract and five insecticides of four different chemical groups were individually added to PDA culture medium, in which the fungus was cultivated. Vegetative growth, sporulation and viability

  19. Host plant affects morphometric variation of Diaphorina citri (Hemiptera: Liviidae).

    Science.gov (United States)

    Paris, Thomson M; Allan, Sandra A; Hall, David G; Hentz, Matthew G; Hetesy, Gabriella; Stansly, Philip A

    2016-01-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is one of the most serious citrus pests worldwide due to its role as vector of huanglongbing or citrus greening disease. While some optimal plant species for ACP oviposition and development have been identified, little is known of the influence of host plants on ACP size and shape. Our goal was to determine how size and shape of ACP wing and body size varies when development occurs on different host plants in a controlled rearing environment. ACP were reared on six different rutaceous species; Bergera koenigii, Citrus aurantifolia, Citrus macrophylla, Citrus maxima, Citrus taiwanica and Murraya paniculata. Adults were examined for morphometric variation using traditional and geometric analysis based on 12 traits or landmarks. ACP reared on C. taiwanica were consistently smaller than those reared on the other plant species. Wing aspect ratio also differed between C. maxima and C. taiwanica. Significant differences in shape were detected with those reared on M. paniculata having narrower wings than those reared on C. macrophylla. This study provides evidence of wing size and shape differences of ACP based on host plant species which potentially may impact dispersal. Further study is needed to determine if behavioral and physiological differences are associated with the observed phenotypic differences.

  20. Host plant affects morphometric variation of Diaphorina citri (Hemiptera: Liviidae

    Directory of Open Access Journals (Sweden)

    Thomson M. Paris

    2016-11-01

    Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri Kuwayama, is one of the most serious citrus pests worldwide due to its role as vector of huanglongbing or citrus greening disease. While some optimal plant species for ACP oviposition and development have been identified, little is known of the influence of host plants on ACP size and shape. Our goal was to determine how size and shape of ACP wing and body size varies when development occurs on different host plants in a controlled rearing environment. ACP were reared on six different rutaceous species; Bergera koenigii, Citrus aurantifolia, Citrus macrophylla, Citrus maxima, Citrus taiwanica and Murraya paniculata. Adults were examined for morphometric variation using traditional and geometric analysis based on 12 traits or landmarks. ACP reared on C. taiwanica were consistently smaller than those reared on the other plant species. Wing aspect ratio also differed between C. maxima and C. taiwanica. Significant differences in shape were detected with those reared on M. paniculata having narrower wings than those reared on C. macrophylla. This study provides evidence of wing size and shape differences of ACP based on host plant species which potentially may impact dispersal. Further study is needed to determine if behavioral and physiological differences are associated with the observed phenotypic differences.

  1. An improved method of DNA extraction from Diaphorina citri for HLB detection

    Science.gov (United States)

    Huanglongbing (HLB) is a devastating disease of citrus that is transmitted by two citrus psyllids. Diaphorina citri transmits Candidatus Liberibacter asiaticus (Las) and Ca. L. americanus (Lam), and Trioza erytreae transmits Ca. L. africanus (Laf). Ca. Liberibacter species can be detected in DNA ex...

  2. Effects of temperature, photoperiod, and rainfall on morphometric variation of Diaphorina citri (Hemiptera: Liviidae)

    Science.gov (United States)

    Phenotypic plasticity provides a mechanism by which an organism can adapt to new or changing environments. Earlier studies have demonstrated the variability of Diaphorina citri Kuwayama (Asian citrus psyllid) population dynamics, but no analysis of morphological changes induced by seasonal or artifi...

  3. Correlation of an electrical penetration graph waveform with walking by Asian citrus psyllid, Diaphorina citri Kuwayama

    Science.gov (United States)

    Electrical penetration graph (EPG) monitoring is being used to study how feeding behavior of the Asian citrus psyllid (ACP) (Diaphorina citri Kuwayama) changes in response to insecticides, in an effort to improve the efficacy of widespread insecticide treatments against the insect in Florida citrus....

  4. Low incidence of Candidatus Liberibacter asiaticus in Diaphorina citri and its host plant Murraya paniculata

    Science.gov (United States)

    Huanglongbing (HLB) is one of the most devastating diseases of citrus worldwide. Candidatus Liberibacter asiaticus is the prevalent species of three HLB-associated Liberibacter species, which is vectored by the psyllid Diaphorina citri. The vector and the bacteria have host plants outside Citrus, ...

  5. Endosymbiont hunting in the metagenome of Asian citrus psyllid (Diaphorina citri) (7th Annual SFAF Meeting, 2012)

    Energy Technology Data Exchange (ETDEWEB)

    Saha, Surya [Cornell University

    2012-06-01

    Surya Saha on "Endosymbiont hunting in the metagenome of Asian citrus psyllid (Diaphorina citri)" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

  6. Repellent Activity of Botanical Oils against Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae).

    Science.gov (United States)

    Kuhns, Emily H; Martini, Xavier; Hoyte, Angel; Stelinski, Lukasz L

    2016-07-14

    The Asian citrus psyllid, Diaphorina citri Kuwayama, is the insect vector of the pathogen causing huanglongbing. We selected three botanical oils to evaluate behavioral activity against D. citri. In laboratory olfactometer assays, fir oil was repellent to D. citri females, while litsea and citronella oils elicited no response from D. citri females. In choice settling experiments, D. citri settled almost completely on control plants rather than on plants treated with fir oil at a 9.5 mg/day release rate. Therefore, we conducted field trials to determine if fir oil reduced D. citri densities in citrus groves. We found no repellency of D. citri from sweet orange resets that were treated with fir oil dispensers releasing 10.4 g/day/tree as compared with control plots. However, we found a two-week decrease in populations of D. citri as compared with controls when the deployment rate of these dispensers was doubled. Our results suggest that treatment of citrus with fir oil may have limited activity as a stand-alone management tool for D. citri and would require integration with other management practices.

  7. Repellent Activity of Botanical Oils against Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae

    Directory of Open Access Journals (Sweden)

    Emily H. Kuhns

    2016-07-01

    Full Text Available The Asian citrus psyllid, Diaphorina citri Kuwayama, is the insect vector of the pathogen causing huanglongbing. We selected three botanical oils to evaluate behavioral activity against D. citri. In laboratory olfactometer assays, fir oil was repellent to D. citri females, while litsea and citronella oils elicited no response from D. citri females. In choice settling experiments, D. citri settled almost completely on control plants rather than on plants treated with fir oil at a 9.5 mg/day release rate. Therefore, we conducted field trials to determine if fir oil reduced D. citri densities in citrus groves. We found no repellency of D. citri from sweet orange resets that were treated with fir oil dispensers releasing 10.4 g/day/tree as compared with control plots. However, we found a two-week decrease in populations of D. citri as compared with controls when the deployment rate of these dispensers was doubled. Our results suggest that treatment of citrus with fir oil may have limited activity as a stand-alone management tool for D. citri and would require integration with other management practices.

  8. Titers of 'Ca. Liberibacter asiaticus' in Murraya paniculata and Murraya-reared Diaphorina citri Are Much Lower than in Citrus and Citrus-reared Psyllids

    National Research Council Canada - National Science Library

    Walter, Abigail; Duan, Y; Hall, David G

    2012-01-01

    Huanglongbing, one of the most devastating diseases of citrus, is associated with the bacterium 'Candidatus Liberibacter asiaticus' vectored by the Asian citrus psyllid, Diaphorina citri, in North America...

  9. Within-orchard edge effects of the azimuth of the sun on Diaphorina citri adults in mature orchards

    Science.gov (United States)

    Huanglongbing (HLB) has been considered the most devastating disease of citrus. The bacterium and vector associated with HLB in Florida are ‘Candidatus Liberibacter asiaticus’ and Diaphorina citri (Asian citrus psyllid), respectively. D. citri is positively phototropic, and higher populations have b...

  10. Diaphorina citri (Hemiptera: Liviidae) responses to microcontroller-buzzer communication signals of potential use in vibration traps

    Science.gov (United States)

    Monitoring of Diaphorina citri Kuwayama populations is an important component of efforts to reduce damage caused by huanglongbing, a devastating disease it vectors in citrus groves. Currently, D. citri is monitored primarily by unbaited sticky traps or visual inspection of trees. A potentially more ...

  11. Spatiotemporal dynamics of the Southern California Asian citrus psyllid (Diaphorina citri) invasion.

    Science.gov (United States)

    Bayles, Brett R; Thomas, Shyam M; Simmons, Gregory S; Grafton-Cardwell, Elizabeth E; Daugherty, Mathew P

    2017-01-01

    Biological invasions are governed by spatial processes that tend to be distributed in non-random ways across landscapes. Characterizing the spatial and temporal heterogeneities of the introduction, establishment, and spread of non-native insect species is a key aspect of effectively managing their geographic expansion. The Asian citrus psyllid (Diaphorina citri), a vector of the bacterium associated with huanglongbing (HLB), poses a serious threat to commercial and residential citrus trees. In 2008, D. citri first began expanding northward from Mexico into parts of Southern California. Using georeferenced D. citri occurrence data from 2008-2014, we sought to better understand the extent of the geographic expansion of this invasive vector species. Our objectives were to: 1) describe the spatial and temporal distribution of D. citri in Southern California, 2) identify the locations of statistically significant D. citri hotspots, and 3) quantify the dynamics of anisotropic spread. We found clear evidence that the spatial and temporal distribution of D. citri in Southern California is non-random. Further, we identified the existence of statistically significant hotspots of D. citri occurrence and described the anisotropic dispersion across the Southern California landscape. For example, the dominant hotspot surrounding Los Angeles showed rapid and strongly asymmetric spread to the south and east. Our study demonstrates the feasibility of quantitative invasive insect risk assessment with the application of a spatial epidemiology framework.

  12. Spatiotemporal dynamics of the Southern California Asian citrus psyllid (Diaphorina citri) invasion

    Science.gov (United States)

    Thomas, Shyam M.; Simmons, Gregory S.; Grafton-Cardwell, Elizabeth E.; Daugherty, Mathew P.

    2017-01-01

    Biological invasions are governed by spatial processes that tend to be distributed in non-random ways across landscapes. Characterizing the spatial and temporal heterogeneities of the introduction, establishment, and spread of non-native insect species is a key aspect of effectively managing their geographic expansion. The Asian citrus psyllid (Diaphorina citri), a vector of the bacterium associated with huanglongbing (HLB), poses a serious threat to commercial and residential citrus trees. In 2008, D. citri first began expanding northward from Mexico into parts of Southern California. Using georeferenced D. citri occurrence data from 2008–2014, we sought to better understand the extent of the geographic expansion of this invasive vector species. Our objectives were to: 1) describe the spatial and temporal distribution of D. citri in Southern California, 2) identify the locations of statistically significant D. citri hotspots, and 3) quantify the dynamics of anisotropic spread. We found clear evidence that the spatial and temporal distribution of D. citri in Southern California is non-random. Further, we identified the existence of statistically significant hotspots of D. citri occurrence and described the anisotropic dispersion across the Southern California landscape. For example, the dominant hotspot surrounding Los Angeles showed rapid and strongly asymmetric spread to the south and east. Our study demonstrates the feasibility of quantitative invasive insect risk assessment with the application of a spatial epidemiology framework. PMID:28278188

  13. Patogenicidade de Beauveria bassiana ao psilídeo Diaphorina citri e compatibilidade do fungo com produtos fitossanitários

    Directory of Open Access Journals (Sweden)

    Ana Paula Ferreira Pinto

    2012-12-01

    Full Text Available O objetivo deste trabalho foi avaliar a patogenicidade de Beauveria bassiana a ninfas de Diaphorina citri (Hemiptera: Psyllidae e verificar a compatibilidade do fungo com produtos fitossanitários e sua persistência em plantas de citros. Ninfas de D. citri foram pulverizadas com B. bassiana, nas concentrações 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 e 1x10(9 conídios mL-1, para determinação da concentração letal. Para avaliação da compatibilidade do fungo com produtos fitossanitários, extrato de nim e cinco inseticidas de quatro grupos químicos diferentes foram incorporados individualmente ao meio de cultura BDA em que o fungo foi cultivado. Avaliaram-se o crescimento vegetativo, a esporulação e a viabilidade do entomopatógeno. Plantas de citros, mantidas em casa de vegetação, foram tratadas primeiramente com os produtos fitossanitários e depois com o entomopatógeno. Avaliaram-se os tempos de exposição de 24 horas e de 7 e 14 dias. O fungo foi patogênico às ninfas de D. citri; a CL50 foi de 0,4x10(7 e a CL90 de 6,7x10(7 conidios mL-1, no décimo dia de avaliação. Em laboratório, os produtos fitosssanitários reduzem o crescimento do fungo. Em casa de vegetação, os produtos não afetam a sobrevivência do fungo nas plantas de citros.

  14. The genetic structure of an invasive pest, the Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae.

    Directory of Open Access Journals (Sweden)

    Aline S Guidolin

    Full Text Available The Asian citrus psyllid Diaphorina citri is currently the major threat to the citrus industry as it is the vector of Candidatus Liberibacter, the causal agent of huanglongbing disease (HLB. D. citri is native to Asia and now colonizes the Americas. Although it has been known in some countries for a long time, invasion routes remain undetermined. There are no efficient control methods for the HLB despite the intensive management tools currently in use. We investigated the genetic variability and structure of populations of D. citri to aid in the decision making processes toward sustainable management of this species/disease. We employed different methods to quantify and compare the genetic diversity and structure of D. citri populations among 36 localities in Brazil, using an almost complete sequence of the cytochrome oxidase I (COI gene. Our analyses led to the identification of two geographically and genetically structured groups. The indices of molecular diversity pointed to a recent population expansion, and we discuss the role of multiple invasion events in this scenario. We also argue that such genetic diversity and population structure may have implications for the best management strategies to be adopted for controlling this psyllid and/or the disease it vectors in Brazil.

  15. Molecular characterization of Wolbachia strains associated with the invasive Asian citrus psyllid Diaphorina citri in Brazil.

    Science.gov (United States)

    Guidolin, A S; Cônsoli, F L

    2013-02-01

    Wolbachia is a symbiont intensively studied due to its ability to interfere with their host's reproduction, and it has been recently proposed as an alternative tool to control insect pests or vectors of diseases. The Asian citrus psyllid Diaphorina citri is an important pest of citrus since it vectors the bacterium that causes the "Huanglongbing" disease in citrus. The frequency and diversity of Wolbachia associated with D. citri is unknown, limiting the utilization of Wolbachia as an alternative strategy for insect management. Thus, we aimed to determine the natural rate of infection, to characterize the Wolbachia strains associated with this psyllid by "multilocus sequencing typing" (MLST) and wsp analysis, and to verify the association of the symbiont to particular genotypes of the host. Analysis indicated Wolbachia infects 100 % of all specimens tested from all 15 sampled populations. MLST revealed the occurrence of five new sequence types (STs) of Wolbachia, while analysis based on the wsp sequences indicated only four different types of Wolbachia. ST-173 was predominant, while the remaining STs were population specific. Analysis of the host-symbiont relationship did not reveal any particular association of Wolbachia and haplotypes or a decrease in nucleotide diversity of D. citri in populations in which more than one ST was recorded. The consequences of the diversity of STs reported are still unknown, but the fact that Wolbachia infection is fixed and that there is one ST with a broad distribution highlights the use of this symbiont as an alternative strategy to control D. citri.

  16. Morphometric comparisons of Diaphorina citri (Hemiptera: Liviidae populations from Iran, USA and Pakistan

    Directory of Open Access Journals (Sweden)

    Mohammadreza Lashkari

    2015-05-01

    Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri Kuwayama (Hemiptera: Liviidae, vector of citrus greening disease pathogen, Huanglongbing (HLB, is considered the most serious pest of citrus in the world. Prior molecular based studies have hypothesized a link between the D. citri in Iran and the USA (Florida. The purpose of this study was to collect morphometric data from D. citri populations from Iran (mtCOI haplotype-1, Florida (mtCOI haplotype-1, and Pakistan (mtCOI haplotype-6, to determine whether different mtCOI haplotypes have a relationship to a specific morphometric variation. 240 samples from 6 ACP populations (Iran—Jiroft, Chabahar; Florida—Ft. Pierce, Palm Beach Gardens, Port St. Lucie; and Pakistan—Punjab were collected for comparison. Measurements of 20 morphological characters were selected, measured and analysed using ANOVA and MANOVA. The results indicate differences among the 6 ACP populations (Wilks’ lambda = 0.0376, F = 7.29, P < 0.0001. The body length (BL, circumanal ring length (CL, antenna length (AL, forewing length (WL and Rs vein length of forewing (RL were the most important characters separating the populations. The cluster analysis showed that the Iran and Florida populations are distinct from each other but separate from the Pakistan population. Thus, three subgroups can be morphologically discriminated within D. citri species in this study, (1 Iran, (2 USA (Florida and (3 Pakistan population. Morphometric comparisons provided further resolution to the mtCOI haplotypes and distinguished the Florida and Iranian populations.

  17. Biocuration and improvement of the Diaphorina citri draft genome assembly with long reads, optical maps and long-range scaffolding

    Science.gov (United States)

    The Asian citrus psyllid (Diaphorina citri Kuwayama) is the insect vector of the bacterium Candidatus Liberibacter asiaticus (CLas), the causal agent for the citrus greening or Huanglongbing disease which threatens citrus industry worldwide. This vector is the primary target of approaches to stop th...

  18. Toxicity of insecticidal soaps to the Asian citrus psyllid (Diaphorina citri) and two of its natural enemies

    Science.gov (United States)

    Insecticidal soaps (hereafter “soaps”) are labeled for use by homeowners and also can be used in citrus grown for the organic market. Soaps control some insect pests and therefore might be an alternative to conventional pesticides for control of the Asian citrus psyllid, Diaphorina citri Kuwayama (...

  19. Analyses of mitogenome sequences revealed that Asian citrus psyllid (Diaphorina citri) from California was related to those from Florida but different from those in Southern China

    Science.gov (United States)

    Asian citrus psyllid (ACP, Diaphorina citri Kuwayama; Hemiptera: Liviidae) transmits “Candidatus Liberibacter asiaticus” (CLas), an unculturable alpha-proteobacterium associated with citrus Huanglongbing (HLB, yellow shoot disease, also called citrus greening disease). HLB is threatening citrus prod...

  20. The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

    Science.gov (United States)

    Stockton, Dara G; Martini, Xavier; Patt, Joseph M; Stelinski, Lukasz L

    2016-01-01

    Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a) whether development on specific host plant species influenced host plant preference in mature D. citri; and b) the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate Rutaceae in the area

  1. Insecticidal suppression of Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae) vector of huanglongbing pathogens.

    Science.gov (United States)

    Qureshi, Jawwad A; Kostyk, Barry C; Stansly, Philip A

    2014-01-01

    Diaphorina citri vectors pathogens that cause 'huanglongbing' or citrus greening disease which poses a serious threat to citrus production worldwide. Vector suppression is critical to reduce disease spread. Efficacy is a main concern when choosing an insecticide. Insecticidal treatments of 49 products or 44 active ingredients (a.i) labeled or experimental were field tested between 2005-2013 as foliar sprays (250 treatments, 39 a.i) or soil applications (47 treatments, 9 a.i) to control D. citri in citrus. A combined effect of nymphal and adult suppression in response to sprays of 23 insecticides representing 9 modes of action (MoA) groups and 3 unknown MoA provided more than 90% reduction of adult D. citri over 24-68 days. Observable effects on nymphs were generally of shorter duration due to rapid maturation of flush. However, reduction of 76-100% nymphs or adults over 99-296 days was seen on young trees receiving drenches of the neonicotinoids imidacloprid, thiamethoxam or clothianidin (MoA 4A) and a novel anthranilic diamide, cyantraniliprole (MoA 28). Effective products identified for foliar sprays to control D. citri provide sufficient MoA groups for rotation to delay evolution of insecticide resistance by D. citri and other pests. However, cyantraniliprole is now the only available alternative for rotation with neonicotinoids in soil application to young trees. Sprays of up to eight of the most effective insecticides could be rotated over a year without repetition of any MoA and little or no recourse to neonicotinoids or cyantraniliprole, so important for protection of young trees. Other considerations effecting decisions of what and when to spray include prevalence of huanglongbing, pest pressure, pre-harvest intervals, overall budget, equipment availability, and conservation of beneficial arthropods. Examples of spray programs utilizing broad-spectrum and relatively selective insecticides are provided to improve vector management and may vary depending on

  2. Multimodal cues drive host-plant assessment in Asian citrus psyllid (Diaphorina citri).

    Science.gov (United States)

    Patt, Joseph M; Meikle, William G; Mafra-Neto, Agenor; Sétamou, Mamoudou; Mangan, Robert; Yang, Chenghai; Malik, Nasir; Adamczyk, John J

    2011-12-01

    Asian citrus psyllid (Diaphorina citri) transmits the causal agent of Huanglongbing, a devastating disease of citrus trees. In this study we measured behavioral responses of D. citri to combinations of visual, olfactory, and gustatory stimuli in test arenas. Stimuli were presented to the psyllids in droplets or lines of an emulsified wax formulation in two different arena types in no-choice tests. First, when placed on a colored ring situated halfway between the center and perimeter of a petri dish, D. citri spent more time on yellow versus gray rings; however, this response disappeared when either gray or yellow wax droplets were applied. When the psyllids were presented with droplets scented with terpenes, the response to both scent and color was increased. The addition of a dilute (≍0.1 M) sucrose solution to the wax droplets increased the magnitude of D. citri responses. Next, groups of D. citri were placed on plastic laboratory film covering a sucrose solution, to mimic a leaf surface. Test stimuli were presented via two 'midribs' made from lines of emulsified wax formulation. Probing levels were measured as a function of color saturation and scent composition, and concentration. The test scents were based on qualitatively major volatiles emitted by Murraya paniculata (L.) Jack, Citrus aurantifolia (Christm.) Swingle, and C. sinensis (L.) Osbeck. The highest probing response was observed on the middle concentration (20-μl scent/10 ml wax formulation) of the C. aurantifolia-scented wax lines. Results indicate that there are interactive effects between the different sensory modalities in directing host-plant assessment behavior.

  3. The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

    Directory of Open Access Journals (Sweden)

    Dara G Stockton

    Full Text Available Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a whether development on specific host plant species influenced host plant preference in mature D. citri; and b the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate

  4. Insecticidal suppression of Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae vector of huanglongbing pathogens.

    Directory of Open Access Journals (Sweden)

    Jawwad A Qureshi

    Full Text Available Diaphorina citri vectors pathogens that cause 'huanglongbing' or citrus greening disease which poses a serious threat to citrus production worldwide. Vector suppression is critical to reduce disease spread. Efficacy is a main concern when choosing an insecticide. Insecticidal treatments of 49 products or 44 active ingredients (a.i labeled or experimental were field tested between 2005-2013 as foliar sprays (250 treatments, 39 a.i or soil applications (47 treatments, 9 a.i to control D. citri in citrus. A combined effect of nymphal and adult suppression in response to sprays of 23 insecticides representing 9 modes of action (MoA groups and 3 unknown MoA provided more than 90% reduction of adult D. citri over 24-68 days. Observable effects on nymphs were generally of shorter duration due to rapid maturation of flush. However, reduction of 76-100% nymphs or adults over 99-296 days was seen on young trees receiving drenches of the neonicotinoids imidacloprid, thiamethoxam or clothianidin (MoA 4A and a novel anthranilic diamide, cyantraniliprole (MoA 28. Effective products identified for foliar sprays to control D. citri provide sufficient MoA groups for rotation to delay evolution of insecticide resistance by D. citri and other pests. However, cyantraniliprole is now the only available alternative for rotation with neonicotinoids in soil application to young trees. Sprays of up to eight of the most effective insecticides could be rotated over a year without repetition of any MoA and little or no recourse to neonicotinoids or cyantraniliprole, so important for protection of young trees. Other considerations effecting decisions of what and when to spray include prevalence of huanglongbing, pest pressure, pre-harvest intervals, overall budget, equipment availability, and conservation of beneficial arthropods. Examples of spray programs utilizing broad-spectrum and relatively selective insecticides are provided to improve vector management and may vary

  5. Efficacy of essential oil of Piper aduncum against nymphs and adults of Diaphorina citri.

    Science.gov (United States)

    Volpe, Haroldo Xl; Fazolin, Murilo; Garcia, Rafael B; Magnani, Rodrigo F; Barbosa, José Carlos; Miranda, Marcelo P

    2016-06-01

    Insecticide application is the main way to control Diaphorina citri. However, it causes environmental contamination, has a negative impact on beneficial organisms and leads to psyllid resistance. The essential oil of Piper aduncum has low toxicity towards the environment and contains dillapiol, which has proven to be effective against several crop pests. Here, we studied its efficacy against nymphs and adults of D. citri under laboratory conditions. Oils with three concentrations of dillapiol (69.3, 79.9 and 85.4%) at 0.5, 0.75 and 1.0% dilutions plus 0.025% adjuvant were tested. All treatments caused 90-100% mortality in nymphs. Topical treatments with oil containing 79.9 and 85.4% dillapiol at 0.75% and 1% dilutions were effective (mortality ≥80%) in adults. However, the essential oil showed no residual activity against adults (mortality ≤30%). Dillapiol-rich oil is a promising compound for D. citri control. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  6. Resistance Management for Asian Citrus Psyllid, Diaphorina citri Kuwayama, in Florida

    Directory of Open Access Journals (Sweden)

    Xue Dong Chen

    2017-09-01

    Full Text Available The Asian citrus psyllid, Diaphorina citri Kuwayma, is one of the most important pests in citrus production. The objective of this study was to evaluate D. citri resistance management with three insecticide rotations and compare them with no rotation and an untreated check. The different insecticides (modes of action tested were: dimethoate, imidacloprid, diflubenzuron, abamectin 3% + thiamethoxam 13.9%, and fenpropathrin. Eggs, nymph, and adult psyllids were counted weekly. Five insecticide applications were made in 2016. Insecticide susceptibility was determined by direct comparison with a laboratory susceptible population and field populations before and after all treatments were applied. Rankings of eggs, nymphs, and adults counted in treated plots were significantly lower than in the untreated control plots after each application. Initially, the resistance ratio (RR50 for each rotation model, as compared with laboratory susceptible strain and the field population before application, was less than 5.76 and 4.31, respectively. However, after five applications with dimethoate, the RR50 using the laboratory and pre-treatment field populations was 42.34 and 34.74, respectively. Our results indicate that effectively rotating modes of action can delay and/or prevent development of insecticide resistance in populations of D. citri.

  7. Living on the Edges: Spatial Niche Occupation of Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae, in Citrus Groves.

    Directory of Open Access Journals (Sweden)

    Mamoudou Sétamou

    Full Text Available The spatial niche occupation of the Asian citrus psyllid, Diaphorina citri Kuwayama, 1908, was evaluated to determine its field colonization and food resource exploitation strategies in citrus groves. Mature grapefruit and sweet orange groves were surveyed as part of an area-wide program in 2009-2010 to determine D. citri population densities and between-tree distribution. In both cultivars, significantly more psyllids were found on perimeter trees throughout the study period suggesting a strong edge effect in D. citri distribution in the groves. D. citri densities and infestation levels gradually declined from the edge to the center of grove. Higher numbers of D. citri were recorded on trees located on the east and south sides of the groves than those on the west and north sides. Citrus groves located at the outer edge of the study with at least one side non-surrounded to other citrus groves harbored significantly more D. citri than groves located within the block cluster and entirely surrounded by other groves. In detailed field studies during 2012, infestation of D. citri started from border trees in the grove where possibly one generation is completed before inner trees become infested. In addition, psyllid densities decreased significantly with increasing distance from the grove edge. Using the selection index, D citri exhibited a strong niche occupation preference for border trees.

  8. Seasonal Population Dynamics of the Asian Citrus Psyllid, Diaphorina citri Kuwayama in Sarawak

    Directory of Open Access Journals (Sweden)

    Stephen L.C. Teck

    2011-01-01

    Full Text Available Problem statement: Effective control of phytophagous pests requires a thorough understanding of their seasonal population dynamics, dispersion behavior, natural enemy activity and climate. To date, although very little detail information had been published on the ecology of Diaphorina citri. The objective of this investigation was to test through field experiment the hypothesis that the major factors influencing local D. citri populations particularly their seasonal population dynamics in Sarawak are (a flushing cycles, (b climate and (c the impact of the primary parasitoids namely Tamarixia radiata and Diaphorencyrtus aligarhensis. Approach: Seasonal abundance D. citri was studied weekly from March 1998 to December 2000 in the 1-ha citrus honey mandarin (Citrus aurantium L. commercial orchard at Jemukan (1° 33'N, 110° 41'E, Kota Samarahan Division, Southwest Sarawak, in Malaysia. Results: Field studies on citrus trees showed that the D. citri population fluctuates throughout the year on citrus honey mandarin in Sarawak. Generations overlapped but adult and egg population peaks for a short period generally coincided with three annual flushing cycles, in August-September, February-March and June-July between March 1998 and December 2000. Conclusion: Psyllid population levels are positively related to the availability of new shoot flushes. Psyllid populations are adversely affected by weather conditions and parasitoids. Adult psyllid populations increased exponentially during periods of flush growth and migration and dispersal of the adults was also related to flushing cycles. Dispersal and colonization of new trees is greatest in September-October, at the onset of the rainy season.

  9. Biology of the Huanglongbing vector Diaphorina citri (Hemiptera: Liviidae) on different host plants.

    Science.gov (United States)

    Alves, G R; Diniz, A J F; Parra, J R P

    2014-04-01

    Although many studies have been conducted on the development and reproductive potential of Diaphorina citri Kuwayama, 1908 (Hemiptera: Liviidae) in different host species, few have evaluated these parameters on different varieties of the same host species. This study evaluated the influence of five commercial varieties of citrus (Citrus spp. L.)--Hamlin, Natal, Pêra, Ponkan, and Valencia-and orange jasmine [Murraya exotica (L.) Jack] on the development of D. citri. Survival rates for the egg stage were highest on orange jasmine (85.7%) and on Valencia (83.3%). The lowest viability of the nymphal stage was also observed on Hamlin, averaging 57.4%. Values for total viability ranged from 65.9 to 32.6%, and were highest on Valencia. The longest egg-adult development time was on Natal, with a mean of 18.4 d; the shortest total development time was on orange jasmine, with a mean of 17.3 d. Based on the fertility life table, the net reproductive rate (Ro) of D. citri was 2.5 times higher when reared on Valencia than on Hamlin. The other parameters (duration of each generation [T], finite rate of increase [lambda], and innate capacity to increase in number [r(m)]) also demonstrated that Valencia is best suited to this insect. The results obtained for the biological parameters and the fertility life table indicate that Valencia and orange jasmine were the most suitable hosts, whereas Hamlin was least suitable for the development of D. citri. These results provide information for the installation of new citrus groves, especially in the choice of varieties to be planted and the location of different varieties within the groves, with a view toward the management of Huanglongbing or HLB.

  10. Susceptibilidad de Diaphorina citri Kuwayama (Hemíptera : Liviidae) a insecticidas en Veracruz, México.

    OpenAIRE

    García Méndez, Víctor Hugo

    2013-01-01

    El uso racional de insecticidas requiere una evaluación previa de la efectividad de los productos para evitar aplicaciones inefectivas o el desarrollo de resistencia. El Psílido Asiático de los Cítricos (PAC), Diaphorina citri es vector del Huanglongbing y requiere estrategias de manejo regional de la resistencia. El objetivo de la investigación fue definir las líneas base de manejo de la resistencia y determinar la susceptibilidad de D. citri a insecticidas convencionales de uso común en aé...

  11. Microbial diversity in Diaphorina citri( Homoptera: Psyllidae) estimated by 16S rDNA analysis using DGGE and RFLP%基于16S rDNA序列的柑桔木虱体内共生菌多样性研究

    Institute of Scientific and Technical Information of China (English)

    殷幼平; 刘婷婷; 田圣超; 胡修峰; 吴东; 王中康

    2011-01-01

    昆虫消化道内是一个复杂的微生态系统,有大量的微生物存在.这些微生物对寄主发育、营养吸收和防御方面都起着重要的作用.本文利用基于16S rRNA基因的PCR-RFLP指纹图谱法和变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)的方法对柑桔黄龙病虫-媒柑桔木虱Diaphorina citri体内细菌菌群多样性进行了研究.经PCR-RFLP分析显示31条序列与变形菌门的假单胞菌科、肠杆菌科、黄单胞菌科、伯克氏菌科、立克次氏菌科和根瘤菌科细菌具有较高同源性.柑桔木虱内共生细菌的优势菌群依次为合胞体共生菌(syncytiumendosymbiont)(同源性99%,5条序列,分离频率31%)、Candidatus Carsonella ruddii和Mycetocyte内共生菌(同源性98%,5条序列,分离频率31%)以及亚洲韧皮杆菌(Candidatus Liberibacter asiaticus)和内共生菌Wolbachia.对柑桔木虱内生细菌16S rDNA V3区序列的PCR-DGGE分析,条带相似性的UPMAGA聚类分析表明,采自九里香Murraya paniculata的柑桔木虱内生细菌大多聚为一支,而来自柑桔的聚在另一支,说明寄主差异对柑桔木虱内生细菌菌群构成的影响大于地理位置的影响.将PCR-DGGE条带测序,序列经GenBank序列比对发现柑桔木虱内生细菌主要属于变形菌门假单胞菌科、立克次氏菌科、肠杆菌科、黄单胞菌科以及厚壁菌门链球菌科和芽孢杆菌科.合胞体共生菌(条带3-4)因其稳定存在于木虱体内且不随柑桔木虱寄主和地理位置的改变而变化,可能是柑桔木虱体内的优势共生菌;内共生细菌Wolbachia也在柑桔木虱内稳定存在,表明我国柑桔木虱感染Wolbachia是普遍现象.PCR-RFLP和PCR-DGGE两种方法相结合较好地反映了柑桔木虱内生细菌菌群的多样性,而且均显示出柑桔木虱内的合胞体共生菌是主要的优势菌群.

  12. Stylet morphometrics and citrus leaf vein structure in relation to feeding behavior of the Asian citrus psyllid Diaphorina citri, vector of citrus huanglongbing bacterium.

    Science.gov (United States)

    Ammar, El-Desouky; Hall, David G; Shatters, Robert G

    2013-01-01

    The Asian citrus psyllid (ACP), Diaphorina citri (Hemiptera: Psyllidae), is the primary vector of the phloem-limited bacterium Candidatus Liberibacter asiaticus (LAS) associated with huanglongbing (HLB, citrus greening), considered the world's most serious disease of citrus. Stylet morphometrics of ACP nymphs and adults were studied in relation to citrus vein structure and to their putative (histologically verified) feeding sites on Valencia orange leaves. ACP nymphs preferred to settle and feed on the lower (abaxial) side of young leaves either on secondary veins or on the sides of the midrib, whereas adults preferred to settle and feed on the upper (adaxial) or lower secondary veins of young or old leaves. Early instar nymphs can reach and probe the phloem probably because the distance to the phloem is considerably shorter in younger than in mature leaves, and is shorter from the sides of the midrib compared to that from the center. Additionally, the thick-walled 'fibrous ring' (sclerenchyma) around the phloem, which may act as a barrier to ACP stylet penetration into the phloem, is more prominent in older than in younger leaves and in the center than on the sides of the midrib. The majority (80-90%) of the salivary sheath termini produced by ACP nymphs and adults that reached a vascular bundle were associated with the phloem, whereas only 10-20% were associated with xylem vessels. Ultrastructural studies on ACP stylets and LAS-infected leaves suggested that the width of the maxillary food canal in first instar nymphs is wide enough for LAS bacteria to traverse during food ingestion (and LAS acquisition). However, the width of the maxillary salivary canal in these nymphs may not be wide enough to accommodate LAS bacteria during salivation (and LAS inoculation) into host plants. This may explain the inability of early instar nymphs to transmit LAS/HLB in earlier reports.

  13. Stylet morphometrics and citrus leaf vein structure in relation to feeding behavior of the Asian citrus psyllid Diaphorina citri, vector of citrus huanglongbing bacterium.

    Directory of Open Access Journals (Sweden)

    El-Desouky Ammar

    Full Text Available The Asian citrus psyllid (ACP, Diaphorina citri (Hemiptera: Psyllidae, is the primary vector of the phloem-limited bacterium Candidatus Liberibacter asiaticus (LAS associated with huanglongbing (HLB, citrus greening, considered the world's most serious disease of citrus. Stylet morphometrics of ACP nymphs and adults were studied in relation to citrus vein structure and to their putative (histologically verified feeding sites on Valencia orange leaves. ACP nymphs preferred to settle and feed on the lower (abaxial side of young leaves either on secondary veins or on the sides of the midrib, whereas adults preferred to settle and feed on the upper (adaxial or lower secondary veins of young or old leaves. Early instar nymphs can reach and probe the phloem probably because the distance to the phloem is considerably shorter in younger than in mature leaves, and is shorter from the sides of the midrib compared to that from the center. Additionally, the thick-walled 'fibrous ring' (sclerenchyma around the phloem, which may act as a barrier to ACP stylet penetration into the phloem, is more prominent in older than in younger leaves and in the center than on the sides of the midrib. The majority (80-90% of the salivary sheath termini produced by ACP nymphs and adults that reached a vascular bundle were associated with the phloem, whereas only 10-20% were associated with xylem vessels. Ultrastructural studies on ACP stylets and LAS-infected leaves suggested that the width of the maxillary food canal in first instar nymphs is wide enough for LAS bacteria to traverse during food ingestion (and LAS acquisition. However, the width of the maxillary salivary canal in these nymphs may not be wide enough to accommodate LAS bacteria during salivation (and LAS inoculation into host plants. This may explain the inability of early instar nymphs to transmit LAS/HLB in earlier reports.

  14. Botanicals, selective insecticides, and predators to control Diaphorina citri (Hemiptera: Liviidae) in citrus orchards.

    Science.gov (United States)

    Khan, Azhar A; Afzal, Muhammad; Qureshi, Jawwad A; Khan, Arif M; Raza, Abubakar M

    2014-12-01

    The Asian citrus psyllid (ACP) Diaphorina citri Kuwayama vectors pathogens that cause huanglongbing (HLB) or citrus greening devastating and economically important disease present in most citrus growing regions. Young citrus shoots are required for psyllid reproduction and development. During winter citrus trees produce little or no new growth. Overwintering adults reproduce in spring on newly emerging shoots also attractive to other pests and beneficial insects. Botanicals and relatively selective insecticides could help to conserve beneficial insects and reduce pest resistance to insecticides. Sprays of Azadirachtin (Neem), Tropane (Datura), Spirotetramat, Spinetoram, and broad-spectrum Imidacloprid were evaluated to control ACP in spring and summer on 10-year-old "Kinow" Citrus reticulata Blanco trees producing new growth. Psyllid populations were high averaging 5-9 nymphs or adults per sample before treatment application. Nymphs or adults were significantly reduced to 0.5-1.5 per sample in all treatments for 3 weeks, average 61%-83% reduction. No significant reduction in ladybeetles Adalia bipunctata, Aneglei scardoni, Cheilomenes sexmaculata, and Coccinella septempunctata was observed. Syrphids, spiders and green lacewings were reduced in treated trees except with Tropane. Studies are warranted to assess impact of these predators on ACP and interaction with insecticides. Observed reduction in ACP populations may not be enough considering its reproductive potential and role in the spread of HLB. Follow-up sprays may be required to achieve additional suppression using rotations of different insecticides. © 2014 Institute of Zoology, Chinese Academy of Sciences.

  15. Species clarification of Isaria isolates used as biocontrol agents against Diaphorina citri (Hemiptera: Liviidae) in Mexico.

    Science.gov (United States)

    Gallou, Adrien; Serna-Domínguez, María G; Berlanga-Padilla, Angélica M; Ayala-Zermeño, Miguel A; Mellín-Rosas, Marco A; Montesinos-Matías, Roberto; Arredondo-Bernal, Hugo C

    2016-03-01

    Entomopathogenic fungi belonging to the genus Isaria (Hypocreales: Cordycipitaceae) are promising candidates for microbial control of insect pests. Currently, the Mexican government is developing a biological control program based on extensive application of Isaria isolates against Diaphorina citri (Hemiptera: Liviidae), a vector of citrus huanglongbing disease. Previous research identified three promising Isaria isolates (CHE-CNRCB 303, 305, and 307; tentatively identified as Isaria fumosorosea) from Mexico. The goal of this work was to obtain a complete morphological and molecular characterization of these isolates. Comparative analysis of morphology established that the isolates showed similar characteristics to Isaria javanica. Multi-gene analysis confirmed the morphological identification by including the three isolates within the I. javanica clade. Additionally, this work demonstrated the misidentifications of three other Isaria isolates (CHE-CNRCB 310 and 324: I. javanica, formerly I. fumosorosea; CHE-CNRCB 393: I. fumosorosea, formerly Isaria farinosa), underlying the need for a full and correct characterization of an isolate before developing a biological control program. Finally, the inter-simple sequence repeat (ISSR) genotyping method revealed that the CHE-CNRCB 303, 305, and 307 isolates belong to three different genotypes. This result indicates that ISSR markers could be used as a tool to monitor their presence in field conditions.

  16. Odorants for surveillance and control of the Asian Citrus Psyllid (Diaphorina citri.

    Directory of Open Access Journals (Sweden)

    Iliano V Coutinho-Abreu

    Full Text Available BACKGROUND: The Asian Citrus Psyllid (ACP, Diaphorina citri, can transmit the bacterium Candidatus Liberibacter while feeding on citrus flush shoots. This bacterium causes Huanglongbing (HLB, a major disease of citrus cultivation worldwide necessitating the development of new tools for ACP surveillance and control. The olfactory system of ACP is sensitive to variety of odorants released by citrus plants and offers an opportunity to develop new attractants and repellents. RESULTS: In this study, we performed single-unit electrophysiology to identify odorants that are strong activators, inhibitors, and prolonged activators of ACP odorant receptor neurons (ORNs. We identified a suite of odorants that activated the ORNs with high specificity and sensitivity, which may be useful in eliciting behavior such as attraction. In separate experiments, we also identified odorants that evoked prolonged ORN responses and antagonistic odorants able to suppress neuronal responses to activators, both of which can be useful in lowering attraction to hosts. In field trials, we tested the electrophysiologically identified activating odorants and identified a 3-odor blend that enhances trap catches by ∼230%. CONCLUSION: These findings provide a set of odorants that can be used to develop affordable and safe odor-based surveillance and masking strategies for this dangerous pest insect.

  17. Influence of Host Plant Species and Flush Growth Stage on the Asian Citrus Psyllid, Diaphorina citri Kuwayama

    Directory of Open Access Journals (Sweden)

    Stephen L.C. Teck

    2011-01-01

    Full Text Available Problem statement: In order to establish the important host range of D. citri, the performance of this psyllid on different rutacous host plants was assessed. Apparent regional differences in host preferences of D. citri as noted above and the need for empirical knowledge of its host preferences in Malaysia where it is lacking. Approach: Experiments on host plant preference between citrus, jasmine orange and curry leaf were conducted over a period of 14 days in the field cage in October 1998 at the agricultural research centre, Semonggok, 15 km from Kuching, Kuching Division, Sarawak in Malaysia. Results: A comparative life cycle study of the Asian Citrus Psyllid Diaphorina citri Kuwayama on three different Rutaceous plant hosts namely citrus, jasmine orange and curry leaf plants was conducted inside the temperature controlled growth chamber maintain at 26 ± 1°C temperature, 75-80% relative humidity with artificial illumination intensity of 10,000 lux equipped with a photoperiod of 14:10 h light: dark regime to permit normal growth. Conclusion: The screenhouse studies confirmed that D. citri can colonise and breed on citrus, jasmine orange and curry leaf plant and that jasmine orange is the preferred host. It is also confirmed that females prefer to oviposit on immature flush 3-10 mm in length. In growth chamber experiments the life cycle of the psyllid was 18.5 days on jasmine orange, 19.0 days on citrus and 23.0 days on curry leaf.

  18. Protein interaction networks at the host–microbe interface in Diaphorina citri, the insect vector of the citrus greening pathogen

    Science.gov (United States)

    Chavez, J. D.; Johnson, R.; Hosseinzadeh, S.; Mahoney, J. E.; Mohr, J. P.; Robison, F.; Zhong, X.; Hall, D. G.; MacCoss, M.; Bruce, J.; Cilia, M.

    2017-01-01

    The Asian citrus psyllid (Diaphorina citri) is the insect vector responsible for the worldwide spread of ‘Candidatus Liberibacter asiaticus’ (CLas), the bacterial pathogen associated with citrus greening disease. Developmental changes in the insect vector impact pathogen transmission, such that D. citri transmission of CLas is more efficient when bacteria are acquired by nymphs when compared with adults. We hypothesize that expression changes in the D. citri immune system and commensal microbiota occur during development and regulate vector competency. In support of this hypothesis, more proteins, with greater fold changes, were differentially expressed in response to CLas in adults when compared with nymphs, including insect proteins involved in bacterial adhesion and immunity. Compared with nymphs, adult insects had a higher titre of CLas and the bacterial endosymbionts Wolbachia, Profftella and Carsonella. All Wolbachia and Profftella proteins differentially expressed between nymphs and adults are upregulated in adults, while most differentially expressed Carsonella proteins are upregulated in nymphs. Discovery of protein interaction networks has broad applicability to the study of host–microbe relationships. Using protein interaction reporter technology, a D. citri haemocyanin protein highly upregulated in response to CLas was found to physically interact with the CLas coenzyme A (CoA) biosynthesis enzyme phosphopantothenoylcysteine synthetase/decarboxylase. CLas pantothenate kinase, which catalyses the rate-limiting step of CoA biosynthesis, was found to interact with a D. citri myosin protein. Two Carsonella enzymes involved in histidine and tryptophan biosynthesis were found to physically interact with D. citri proteins. These co-evolved protein interaction networks at the host–microbe interface are highly specific targets for controlling the insect vector responsible for the spread of citrus greening. PMID:28386418

  19. Characterization of a recombinant Cathepsin B-Like cysteine peptidase from Diaphorina citri Kuwayama (Hemiptera: Liviidae): A putative target control of citrus huanglongbing

    Science.gov (United States)

    Huanglongbing (HLB) is one of the most destructive disease affecting citrus plants. The causal agent is associated with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) spread by the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae). Among the control strategies for H...

  20. Titers of 'Ca. Liberibacter asiaticus' in Murraya paniculata and Murraya-reared Diaphorina citri are much lower than in citrus and citrus-reared psyllids

    Science.gov (United States)

    Huanglongbing, one of the most devastating diseases of citrus, is associated with the bacterium Candidatus Liberibacter asiaticus vectored by the Asian citrus psyllid, Diaphorina citri, in North America. Murraya paniculata is a common ornamental plant that is an alternate host of Ca. L. asiaticus an...

  1. Cloning and expressing a highly functional and substrate specific farnesoic acid o-methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama)

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri, transmits a phloem-limited bacterium, Candidatus Liberibacter asiaticus that causes citrus greening disease. Because juvenile hormone (JH) plays an important role in adult and nymphal development, we studied the final steps in juvenile hormone biosynthesis...

  2. Antennal and Abdominal Transcriptomes Reveal Chemosensory Genes in the Asian Citrus Psyllid, Diaphorina citri.

    Directory of Open Access Journals (Sweden)

    Zhongzhen Wu

    Full Text Available The Asian citrus psyllid, Diaphorina citri is the principal vector of the highly destructive citrus disease called Huanglongbing (HLB or citrus greening, which is a major threat to citrus cultivation worldwide. More effective pest control strategies against this pest entail the identification of potential chemosensory proteins that could be used in the development of attractants or repellents. However, the molecular basis of olfaction in the Asian citrus psyllid is not completely understood. Therefore, we performed this study to analyze the antennal and abdominal transcriptome of the Asian citrus psyllid. We identified a large number of transcripts belonging to nine chemoreception-related gene families and compared their expression in male and female adult antennae and terminal abdomen. In total, 9 odorant binding proteins (OBPs, 12 chemosensory proteins (CSPs, 46 odorant receptors (ORs, 20 gustatory receptors (GRs, 35 ionotropic receptors (IRs, 4 sensory neuron membrane proteins (SNMPs and 4 different gene families encoding odorant-degrading enzymes (ODEs: 80 cytochrome P450s (CYPs, 12 esterase (ESTs, and 5 aldehyde dehydrogenases (ADE were annotated in the D. citri antennal and abdominal transcriptomes. Our results revealed that a large proportion of chemosensory genes exhibited no distinct differences in their expression patterns in the antennae and terminal abdominal tissues. Notably, RNA sequencing (RNA-seq data and quantitative real time-PCR (qPCR analyses showed that 4 DictOBPs, 4 DictCSPs, 4 DictIRs, 1 DictSNMP, and 2 DictCYPs were upregulated in the antennae relative to that in terminal abdominal tissues. Furthermore, 2 DictOBPs (DictOBP8 and DictOBP9, 2 DictCSPs (DictOBP8 and DictOBP12, 4 DictIRs (DictIR3, DictIR6, DictIR10, and DictIR35, and 1 DictCYP (DictCYP57 were expressed at higher levels in the male antennae than in the female antennae. Our study provides the first insights into the molecular basis of chemoreception in this

  3. β-caryophyllene emitted from a transgenic Arabidopsis or chemical dispenser repels Diaphorina citri, vector of Candidatus Liberibacters.

    Science.gov (United States)

    Alquézar, Berta; Volpe, Haroldo Xavier Linhares; Magnani, Rodrigo Facchini; de Miranda, Marcelo Pedreira; Santos, Mateus Almeida; Wulff, Nelson Arno; Bento, Jose Mauricio Simões; Parra, José Roberto Postali; Bouwmeester, Harro; Peña, Leandro

    2017-07-17

    Production of citrus, the main fruit tree crop worldwide, is severely threatened by Huanglongbing (HLB), for which as yet a cure is not available. Spread of this bacterial disease in America and Asia is intimately connected with dispersal and feeding of the insect vector Diaphorina citri, oligophagous on rutaceous host plants. Effective control of this psyllid is an important component in successful HLB management programs. Volatiles released from the non-host guava have been shown to be repellent to the psyllid and to inhibit its response to citrus odour. By analysing VOC emission from guava we identified one volatile compound, (E)-β-caryophyllene, which at certain doses exerts a repellent effect on D. citri. Non-host plant rejection mediated by (E)-β-caryophyllene is demonstrated here by using Arabidopsis over-expression and knock-out lines. For the first time, results indicate that genetically engineered Arabidopsis plants with modified emission of VOCs can alter the behaviour of D. citri. This study shows that transgenic plants with an inherent ability to release (E)-β-caryophyllene can potentially be used in new protection strategies of citrus trees against HLB.

  4. Fluctuación poblacional de Diaphorina citri Kuwayana en limón persa (Citrus latifolia Tanaka) en la sabana de Huimanguillo, Tabasco.

    OpenAIRE

    García Garduza, Diógenes

    2012-01-01

    Diaphorina citri Kuwayama es el vector de la enfermedad Huanglongbing, la cual se encuentra actualmente en 14 de los 23 estados productores de cítricos en México. El estado de Tabasco, donde se cultivan 15, 406 ha de estos frutales, aún no es afectado por la enfermedad, por lo cual es importante tomar medidas para controlar las poblaciones del vector, cuya presencia en dicho estado fue detectada en 2005. El objetivo de este trabajo fue determinar la fluctuación poblacional de D. citri y la r...

  5. Detection of 'Candidatus Liberibacter asiaticus' in Diaphorina citri and its importance in the management of citrus huanglongbing in Florida.

    Science.gov (United States)

    Manjunath, K L; Halbert, S E; Ramadugu, C; Webb, S; Lee, R F

    2008-04-01

    Citrus huanglongbing (HLB or citrus greening), is a highly destructive disease that has been spreading in both Florida and Brazil. Its psyllid vector, Diaphorina citri Kuwayama, has spread to Texas and Mexico, thus threatening the future of citrus production elsewhere in mainland North America. Even though sensitive diagnostic methods have been developed for detection of the causal organisms, Candidatus Liberibacter spp., the pathogen cannot be detected consistently in plants until symptoms develop, presumably because of low titer and uneven distribution of the causal bacteria in nonsymptomatic tissues. In the present study, TaqMan based real-time quantitative polymerase chain reaction methodology was developed for detection of 'Ca. L. asiaticus' in D. citri. Over 1,200 samples of psyllid adults and nymphs, collected from various locations in Florida, from visually healthy and HLB symptomatic trees at different times of the year were analyzed to monitor the incidence and spread of HLB. The results showed that spread of 'Ca. L. asiaticus' in an area may be detected one to several years before the development of HLB symptoms in plants. The study suggests that discount garden centers and retail nurseries may have played a significant role in the widespread distribution of psyllids and plants carrying HLB pathogens in Florida.

  6. Characterization of a Recombinant Cathepsin B-Like Cysteine Peptidase from Diaphorina citri Kuwayama (Hemiptera: Liviidae): A Putative Target for Control of Citrus Huanglongbing.

    Science.gov (United States)

    Ferrara, Taíse Fernanda da Silva; Schneider, Vanessa Karine; Kishi, Luciano Takeshi; Carmona, Adriana Karaoglanovic; Alves, Marcio Fernando Madureira; Belasque-Júnior, Jose; Rosa, José César; Hunter, Wayne Brian; Henrique-Silva, Flávio; Soares-Costa, Andrea

    2015-01-01

    Huanglonbing (HLB) is one of the most destructive disease affecting citrus plants. The causal agent is associated with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) and the psyllid Diaphorina citri, vector of disease, that transmits the bacterium associated with HLB. The control of disease can be achieved by suppressing either the bacterium or the vector. Among the control strategies for HLB disease, one of the widely used consists in controlling the enzymes of the disease vector, Diaphorina citri. The insect Diaphorina citri belongs to the order Hemiptera, which frequently have cysteine peptidases in the gut. The importance of this class of enzymes led us to search for enzymes in the D. citri transcriptome for the establishment of alternatives strategies for HLB control. In this study, we reported the identification and characterization of a cathepsin B-like cysteine peptidase from D. citri (DCcathB). DCcathB was recombinantly expressed in Pichia pastoris, presenting a molecular mass of approximately 50 kDa. The enzyme hydrolyzed the fluorogenic substrate Z-F-R-AMC (Km = 23.5 μM) and the selective substrate for cathepsin B, Z-R-R-AMC (Km = 6.13 μM). The recombinant enzyme was inhibited by the cysteine protease inhibitors E64 (IC50 = 0.014 μM) and CaneCPI-4 (Ki = 0.05 nM) and by the selective cathepsin B inhibitor CA-074 (IC50 = 0.095 nM). RT-qPCR analysis revealed that the expression of the DCcathB in nymph and adult was approximately 9-fold greater than in egg. Moreover, the expression of this enzyme in the gut was 175-fold and 3333-fold higher than in the remaining tissues and in the head, respectively, suggesting that DCcathB can be a target for HLB control.

  7. Characterization of a Recombinant Cathepsin B-Like Cysteine Peptidase from Diaphorina citri Kuwayama (Hemiptera: Liviidae: A Putative Target for Control of Citrus Huanglongbing.

    Directory of Open Access Journals (Sweden)

    Taíse Fernanda da Silva Ferrara

    Full Text Available Huanglonbing (HLB is one of the most destructive disease affecting citrus plants. The causal agent is associated with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas and the psyllid Diaphorina citri, vector of disease, that transmits the bacterium associated with HLB. The control of disease can be achieved by suppressing either the bacterium or the vector. Among the control strategies for HLB disease, one of the widely used consists in controlling the enzymes of the disease vector, Diaphorina citri. The insect Diaphorina citri belongs to the order Hemiptera, which frequently have cysteine peptidases in the gut. The importance of this class of enzymes led us to search for enzymes in the D. citri transcriptome for the establishment of alternatives strategies for HLB control. In this study, we reported the identification and characterization of a cathepsin B-like cysteine peptidase from D. citri (DCcathB. DCcathB was recombinantly expressed in Pichia pastoris, presenting a molecular mass of approximately 50 kDa. The enzyme hydrolyzed the fluorogenic substrate Z-F-R-AMC (Km = 23.5 μM and the selective substrate for cathepsin B, Z-R-R-AMC (Km = 6.13 μM. The recombinant enzyme was inhibited by the cysteine protease inhibitors E64 (IC50 = 0.014 μM and CaneCPI-4 (Ki = 0.05 nM and by the selective cathepsin B inhibitor CA-074 (IC50 = 0.095 nM. RT-qPCR analysis revealed that the expression of the DCcathB in nymph and adult was approximately 9-fold greater than in egg. Moreover, the expression of this enzyme in the gut was 175-fold and 3333-fold higher than in the remaining tissues and in the head, respectively, suggesting that DCcathB can be a target for HLB control.

  8. Metabolomic Response to Huanglongbing: Role of Carboxylic Compounds in Citrus sinensis Response to 'Candidatus Liberibacter asiaticus' and Its Vector, Diaphorina citri.

    Science.gov (United States)

    Killiny, Nabil; Nehela, Yasser

    2017-08-01

    Huanglongbing, a destructive disease of citrus, is caused by the fastidious bacterium 'Candidatus Liberibacter asiaticus' and transmitted by Asian citrus psyllid, Diaphorina citri. The impact of 'Ca. L. asiaticus' infection or D. citri infestation on Valencia sweet orange (Citrus sinensis) leaf metabolites was investigated using gas chromatography mass spectrometry, followed by gene expression analysis for 37 genes involved in jasmonic acid (JA), salicylic acid (SA), and proline-glutamine pathways. The total amino acid abundance increased after 'Ca. L. asiaticus' infection, while the total fatty acids increased dramatically after infestation with D. citri, compared with control plants. Seven amino acids (glycine, l-isoleucine, l-phenylalanine, l-proline, l-serine, l-threonine, and l-tryptophan) and five organic acids (benzoic acid, citric acid, fumaric acid, SA, and succinic acid) increased in 'Ca. L. asiaticus'-infected plants. On the other hand, the abundance of trans-JA and its precursor α-linolenic increased in D. citri-infested plants. Surprisingly, the double attack of both D. citri infestation and 'Ca. L. asiaticus' infection moderated the metabolic changes in all chemical classes studied. In addition, the gene expression analysis supported these results. Based on these findings, we suggest that, although amino acids such as phenylalanine are involved in citrus defense against 'Ca. L. asiaticus' infection through the activation of an SA-mediated pathway, fatty acids, especially α-linolenic acid, are involved in defense against D. citri infestation via the induction of a JA-mediated pathway.

  9. Citrus tristeza virus-based RNAi in citrus plants induces gene silencing in Diaphorina citri, a phloem-sap sucking insect vector of citrus greening disease (Huanglongbing).

    Science.gov (United States)

    Hajeri, Subhas; Killiny, Nabil; El-Mohtar, Choaa; Dawson, William O; Gowda, Siddarame

    2014-04-20

    A transient expression vector based on Citrus tristeza virus (CTV) is unusually stable. Because of its stability it is being considered for use in the field to control Huanglongbing (HLB), which is caused by Candidatus Liberibacter asiaticus (CLas) and vectored by Asian citrus psyllid, Diaphorina citri. In the absence of effective control strategies for CLas, emphasis has been on control of D. citri. Coincident cohabitation in phloem tissue by CLas, D. citri and CTV was exploited to develop a novel method to mitigate HLB through RNA interference (RNAi). Since CTV has three RNA silencing suppressors, it was not known if CTV-based vector could induce RNAi in citrus. Yet, expression of sequences targeting citrus phytoene desaturase gene by CTV-RNAi resulted in photo-bleaching phenotype. CTV-RNAi vector, engineered with truncated abnormal wing disc (Awd) gene of D. citri, induced altered Awd expression when silencing triggers ingested by feeding D. citri nymphs. Decreased Awd in nymphs resulted in malformed-wing phenotype in adults and increased adult mortality. This impaired ability of D. citri to fly would potentially limit the successful vectoring of CLas bacteria between citrus trees in the grove. CTV-RNAi vector would be relevant for fast-track screening of candidate sequences for RNAi-mediated pest control. Copyright © 2014. Published by Elsevier B.V.

  10. Perspectives to the use of entomopathogenic fungi for biological control of Diaphorina citri in Mexico

    OpenAIRE

    Sánchez-Peña, S. R.; Guizar-Guzmán, L.; Torres-Acosta, R. I.; López-Arroyo, J. I.; Casique-Valdés, R.

    2014-01-01

    Entomopathogenic fungi are natural enemies of D. citri adults in Mexico. Natural prevalence (%) of fungi on live adults in central Veracruz (summer) and southern Tamaulipas (fall) were: Hirsutella citriformis (7-35); Isaria fumosorosea (5-15); Lecanicillium (2 in Tamaulipas), and Beauveria (

  11. Diaphorina citri Induces Huanglongbing-Infected Citrus Plant Volatiles to Repel and Reduce the Performance of Propylaea japonica

    Science.gov (United States)

    Lin, Yongwen; Lin, Sheng; Akutse, Komivi S.; Hussain, Mubasher; Wang, Liande

    2016-01-01

    Transmission of plant pathogens through insect vectors is a complex biological process involving interactions between the host plants, insects, and pathogens. Simultaneous impact of the insect damage and pathogenic bacteria in infected host plants induce volatiles that modify not only the behavior of its insect vector but also of their natural enemies, such as parasitoid wasps. Therefore, it is essential to understand how insects such as the predator ladybird beetle responds to volatiles emitted from a host plant and how the disease transmission alters the interactions between predators, vector, pathogens, and plants. In this study, we investigated the response of Propylaea japonica to volatiles from citrus plants damaged by Diaphorina citri and Candidatus Liberibacter asiaticus through olfactometer bioassays. Synthetic chemical blends were also used to determine the active compounds in the plant volatile. The results showed that volatiles emitted by healthy plants attracted more P. japonica than other treatments, due to the presence of high quantities of D-limonene and beta-ocimene, and the lack of methyl salicylate. When using synthetic chemicals in the olfactory tests, we found that D-limonene attracted P. japonica while methyl salicylate repelled the predator. However, beta-ocimene attracted the insects at lower concentrations but repelled them at higher concentrations. These results indicate that P. japonica could not efficiently search for its host by using volatile cues emitted from psyllids- and Las bacteria-infected citrus plants. PMID:28083006

  12. Diaphorina citri Induces Huanglongbing-Infected Citrus Plant Volatiles to Repel and Reduce the Performance of Propylaea japonica.

    Science.gov (United States)

    Lin, Yongwen; Lin, Sheng; Akutse, Komivi S; Hussain, Mubasher; Wang, Liande

    2016-01-01

    Transmission of plant pathogens through insect vectors is a complex biological process involving interactions between the host plants, insects, and pathogens. Simultaneous impact of the insect damage and pathogenic bacteria in infected host plants induce volatiles that modify not only the behavior of its insect vector but also of their natural enemies, such as parasitoid wasps. Therefore, it is essential to understand how insects such as the predator ladybird beetle responds to volatiles emitted from a host plant and how the disease transmission alters the interactions between predators, vector, pathogens, and plants. In this study, we investigated the response of Propylaea japonica to volatiles from citrus plants damaged by Diaphorina citri and Candidatus Liberibacter asiaticus through olfactometer bioassays. Synthetic chemical blends were also used to determine the active compounds in the plant volatile. The results showed that volatiles emitted by healthy plants attracted more P. japonica than other treatments, due to the presence of high quantities of D-limonene and beta-ocimene, and the lack of methyl salicylate. When using synthetic chemicals in the olfactory tests, we found that D-limonene attracted P. japonica while methyl salicylate repelled the predator. However, beta-ocimene attracted the insects at lower concentrations but repelled them at higher concentrations. These results indicate that P. japonica could not efficiently search for its host by using volatile cues emitted from psyllids- and Las bacteria-infected citrus plants.

  13. Oral delivery of double-stranded RNAs induces mortality in nymphs and adults of the Asian citrus psyllid, Diaphorina citri

    Science.gov (United States)

    Galdeano, Diogo Manzano; Breton, Michèle Claire; Lopes, João Roberto Spotti; Falk, Bryce W.

    2017-01-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is one of the most important citrus pests. ACP is the vector of the phloem-limited bacteria Candidatus Liberibacter americanus and Candidatus Liberibacter asiaticus, the causal agents of the devastating citrus disease huanglongbing (HLB). The management of HLB is based on the use of healthy young plants, eradication of infected plants and chemical control of the vector. RNA interference (RNAi) has proven to be a promising tool to control pests and explore gene functions. Recently, studies have reported that target mRNA knockdown in many insects can be induced through feeding with double-stranded RNA (dsRNA). In the current study, we targeted the cathepsin D, chitin synthase and inhibitor of apoptosis genes of adult and nymph ACP by feeding artificial diets mixed with dsRNAs and Murraya paniculata leaves placed in dsRNAs solutions, respectively. Adult ACP mortality was positively correlated with the amount of dsRNA used. Both nymphs and adult ACP fed dsRNAs exhibited significantly increased mortality over time compared with that of the controls. Moreover, qRT-PCR analysis confirmed the dsRNA-mediated RNAi effects on target mRNAs. These results showed that RNAi can be a powerful tool for gene function studies in ACP and perhaps for HLB control. PMID:28282380

  14. Two-Spotted Ladybeetle Adalia bipunctata L. (Coleoptera: Coccinellidae): A Commercially Available Predator to Control Asian Citrus Psyllid Diaphorina citri (Hemiptera: Liviidae)

    Science.gov (United States)

    Afzal, Muhammad; Stansly, Philip A.

    2016-01-01

    The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae) is an economically important pest of citrus because it serves as a vector of the causal pathogens of huanglongbing (HLB) also known as citrus greening disease. The increased use of insecticides for control of D. citri negatively impacts several natural enemies including some effective ladybeetle species which are not available commercially. The two-spotted ladybeetle, Adalia bipunctata (Coleoptera: Coccinellidae) is found in some crop and forest ecosystems of Asia, Europe and North America and available commercially. It is known to attack aphids and mealybugs but there are no published records of feeding on psyllids. We evaluated suitability and preference of A. bipunctata for nymphs of D. citri compared to corn leaf aphid Rhopalosiphum maidis (Hemiptera: Aphididae) a global pest of cereal crops and prey for many predaceous insects. We also compared development and reproduction of A. bipunctata on these two species with frozen eggs of the Mediterranean flour moth Ephestia kuehniella (Lepidoptera: Pyralidae) at 25°C. Initially, more D. citri than R. maidis nymphs were consumed in the no-choice tests although final consumption by larva and adult of A. bipunctata did not differ in the choice and no-choice tests. Larval development was prolonged by one day on D. citri compared to R. maidis nymphs but did not differ between either of these diets and E. kuehniella. Larval survival to adult averaged 93–100% and was not impacted by diet. Adult life span did not differ between diets although those on D. citri and R. maidis nymphs weighed less and produced fewer but more fertile eggs than on E. kuehniella eggs. Significant reduction of D. citri nymphs averaging 54% was observed in colonies caged with adult A. bipunctata on field planted citrus. R° (net reproductive rate) was least for beetles fed R. maidis, but otherwise there were no significant differences in demographic parameters. Successful

  15. Two-Spotted Ladybeetle Adalia bipunctata L. (Coleoptera: Coccinellidae): A Commercially Available Predator to Control Asian Citrus Psyllid Diaphorina citri (Hemiptera: Liviidae).

    Science.gov (United States)

    Khan, Azhar A; Qureshi, Jawwad A; Afzal, Muhammad; Stansly, Philip A

    2016-01-01

    The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae) is an economically important pest of citrus because it serves as a vector of the causal pathogens of huanglongbing (HLB) also known as citrus greening disease. The increased use of insecticides for control of D. citri negatively impacts several natural enemies including some effective ladybeetle species which are not available commercially. The two-spotted ladybeetle, Adalia bipunctata (Coleoptera: Coccinellidae) is found in some crop and forest ecosystems of Asia, Europe and North America and available commercially. It is known to attack aphids and mealybugs but there are no published records of feeding on psyllids. We evaluated suitability and preference of A. bipunctata for nymphs of D. citri compared to corn leaf aphid Rhopalosiphum maidis (Hemiptera: Aphididae) a global pest of cereal crops and prey for many predaceous insects. We also compared development and reproduction of A. bipunctata on these two species with frozen eggs of the Mediterranean flour moth Ephestia kuehniella (Lepidoptera: Pyralidae) at 25°C. Initially, more D. citri than R. maidis nymphs were consumed in the no-choice tests although final consumption by larva and adult of A. bipunctata did not differ in the choice and no-choice tests. Larval development was prolonged by one day on D. citri compared to R. maidis nymphs but did not differ between either of these diets and E. kuehniella. Larval survival to adult averaged 93-100% and was not impacted by diet. Adult life span did not differ between diets although those on D. citri and R. maidis nymphs weighed less and produced fewer but more fertile eggs than on E. kuehniella eggs. Significant reduction of D. citri nymphs averaging 54% was observed in colonies caged with adult A. bipunctata on field planted citrus. R° (net reproductive rate) was least for beetles fed R. maidis, but otherwise there were no significant differences in demographic parameters. Successful

  16. Risk assessment of various insecticides used for management of Asian citrus psyllid, Diaphorina citri in Florida citrus, against honey bee, Apis mellifera.

    Science.gov (United States)

    Chen, Xue Dong; Gill, Torrence A; Pelz-Stelinski, Kirsten S; Stelinski, Lukasz L

    2017-01-23

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a major pest of citrus trees worldwide. A wide variety of insecticides are used to manage D. citri populations within citrus groves in Florida. However, in areas shared by citrus growers and beekeepers the use of insecticides may increase the risks of Apis mellifera  L. (Hymenoptera: Apidae) loss and contaminated honey. The objective of this research was to determine the environmental toxicity of insecticides, spanning five different modes of action used to control D. citri, to A. mellifera. The insecticides investigated were imidacloprid, fenpropathrin, dimethoate, spinetoram and diflubenzuron. In laboratory experiments, LD50 values were determined and ranged from 0.10 to 0.53 ng/μl for imidacloprid, fenpropathrin, dimethoate and spinetoram. LD50 values for diflubenzuron were >1000 ng/μl. Also, a hazard quotient was determined and ranged from 1130.43 to 10893.27 for imidacloprid, fenpropathrin, dimethoate, and spinetoram. This quotient was mellifera 3 and 7 days after application. Spinetoram and imidacloprid were moderately toxic to A. mellifera at the recommended rates for D. citri. Diflubenzuron was not toxic to A. mellifera in the field as compared with untreated control plots. Phenoloxidase (PO) activity of A. mellifera was higher than in untreated controls when A. mellifera were exposed to 14 days old residues. The results indicate that diflubenzuron may be safe to apply in citrus when A. mellifera are foraging, while most insecticides used for management of D. citri in citrus are likely hazardous under various exposure scenarios.

  17. Impacts of Horticultural Mineral Oils and Two Insecticide Practices on Population Fluctuation of Diaphorina citri and Spread of Huanglongbing in a Citrus Orchard in Sarawak

    Directory of Open Access Journals (Sweden)

    Stephen Chan Teck Leong

    2012-01-01

    Full Text Available Aspects of the incidence and spread of the citrus disease huanglongbing (HLB in relation to the vector Diaphorina citri population fluctuation were studied from January 1999 to December 2001 seasons in a 0.8 ha citrus orchard at Jemukan (1° 33′N, 110° 41′E, Southwest Sarawak in Malaysia. In relation to insecticide and horticultural mineral oils (HMOs use, levels of HLB infection rose quite rapidly over the next 3 years in the unsprayed control and less rapidly in the other treatments such as imidacloprid, nC24HMO, and triazophos/cypermethrin/chlorpyrifos. Levels of HLB as determined by Polymerase Chain Reaction (PCR were 42.2%, 9.4%, 11.4%, and 22.7%, respectively. The effects of nC24HMO and conventional pesticides on the citrus psyllid population and parasitoids in citrus orchard were also determined.

  18. Impacts of horticultural mineral oils and two insecticide practices on population fluctuation of Diaphorina citri and spread of Huanglongbing in a citrus orchard in Sarawak.

    Science.gov (United States)

    Leong, Stephen Chan Teck; Abang, Fatimah; Beattie, Andrew; Kueh, Roland Jui Heng; Wong, Sing King

    2012-01-01

    Aspects of the incidence and spread of the citrus disease huanglongbing (HLB) in relation to the vector Diaphorina citri population fluctuation were studied from January 1999 to December 2001 seasons in a 0.8 ha citrus orchard at Jemukan (1° 33'N, 110° 41'E), Southwest Sarawak in Malaysia. In relation to insecticide and horticultural mineral oils (HMOs) use, levels of HLB infection rose quite rapidly over the next 3 years in the unsprayed control and less rapidly in the other treatments such as imidacloprid, nC24HMO, and triazophos/cypermethrin/chlorpyrifos. Levels of HLB as determined by Polymerase Chain Reaction (PCR) were 42.2%, 9.4%, 11.4%, and 22.7%, respectively. The effects of nC(24)HMO and conventional pesticides on the citrus psyllid population and parasitoids in citrus orchard were also determined.

  19. Innate and Conditioned Responses to Chemosensory and Visual Cues in Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae), Vector of Huanglongbing Pathogens.

    Science.gov (United States)

    Patt, Joseph M; Stockton, Dara; Meikle, William G; Sétamou, Mamoudou; Mafra-Neto, Agenor; Adamczyk, John J

    2014-11-19

    Asian citrus psyllid (Diaphorina citri) transmits Huanglongbing, a devastating disease that threatens citrus trees worldwide. A better understanding of the psyllid's host-plant selection process may lead to the development of more efficient means of monitoring it and predicting its movements. Since behavioral adaptations, such as associative learning, may facilitate recognition of suitable host-plants, we examined whether adult D. citri could be conditioned to visual and chemosensory stimuli from host and non-host-plant sources. Response was measured as the frequency of salivary sheaths, the residue of psyllid probing activity, in a line of emulsified wax on the surface of a test arena. The psyllids displayed both appetitive and aversive conditioning to two different chemosensory stimuli. They could also be conditioned to recognize a blue-colored probing substrate and their response to neutral visual cues was enhanced by chemosensory stimuli. Conditioned psyllids were sensitive to the proportion of chemosensory components present in binary mixtures. Naïve psyllids displayed strong to moderate innate biases to several of the test compounds. While innate responses are probably the psyllid's primary behavioral mechanism for selecting host-plants, conditioning may enhance its ability to select host-plants during seasonal transitions and dispersal.

  20. Innate and Conditioned Responses to Chemosensory and Visual Cues in Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae, Vector of Huanglongbing Pathogens

    Directory of Open Access Journals (Sweden)

    Joseph M. Patt

    2014-11-01

    Full Text Available Asian citrus psyllid (Diaphorina citri transmits Huanglongbing, a devastating disease that threatens citrus trees worldwide. A better understanding of the psyllid’s host-plant selection process may lead to the development of more efficient means of monitoring it and predicting its movements. Since behavioral adaptations, such as associative learning, may facilitate recognition of suitable host-plants, we examined whether adult D. citri could be conditioned to visual and chemosensory stimuli from host and non-host-plant sources. Response was measured as the frequency of salivary sheaths, the residue of psyllid probing activity, in a line of emulsified wax on the surface of a test arena. The psyllids displayed both appetitive and aversive conditioning to two different chemosensory stimuli. They could also be conditioned to recognize a blue-colored probing substrate and their response to neutral visual cues was enhanced by chemosensory stimuli. Conditioned psyllids were sensitive to the proportion of chemosensory components present in binary mixtures. Naïve psyllids displayed strong to moderate innate biases to several of the test compounds. While innate responses are probably the psyllid’s primary behavioral mechanism for selecting host-plants, conditioning may enhance its ability to select host-plants during seasonal transitions and dispersal.

  1. DIVERSIDAD GENÉTICA DE Diaphorina citri EN CULTIVOS CÍTRICOS DEL VALLE DEL CAUCA Y QUINDÍO (COLOMBIA

    Directory of Open Access Journals (Sweden)

    MIGUEL ANGEL MONCAYO-DONOSO

    Full Text Available El psílido asiático Diaphorina citri es el principal vector de la bacteria Candidatus Liberibacter que causa la enfermedad Huanglongbing, considerada devastadora en la citricultura mundial y aún no reportada en Colombia. La variabilidad genética poblacional de D. citri fue estudiadautilizando la técnica de secuenciación del gen COI mitocondrial como marcador molecular. Se realizaron colectas de adultos en zonas productoras de cítricos del Valle del Cauca y el Quindío. La amplificación se realizó con dos parejas de cebadores específicos para hemípteros. Los productos de la PCR fueron secuenciados en Macrogen-Korea obteniéndose un total de 124 secuencias. En el análisis bioinformático se utilizaron los programas Vector NTI 11.5, Arlequín V 3.5, MEGA 5 y MAFFT 6. Los índices de diversidad molecular fueron similares entre poblaciones, mostrando un mismo origen, separación reciente de poblaciones y que no existe diferenciación genética significativa que pueda albergar variantes de la bacteria; sin embargo, el índice de diversidad haplotípica fue más alto que el de diversidad nucleotídica y este último índice presentó bajo número de sitios polimórficos, indicando que las poblaciones de D. citri están en expansión. El estudio de la variabilidad genética del vector representa una herramienta para predecir escenarios probables de propagación de la enfermedad.

  2. Essential Oil Variation from Twenty Two Genotypes of Citrus in Brazil-Chemometric Approach and Repellency Against Diaphorina citri Kuwayama.

    Science.gov (United States)

    Andrade, Moacir Dos Santos; Ribeiro, Leandro do Prado; Borgoni, Paulo Cesar; Silva, Maria Fátima das Graças Fernandes da; Forim, Moacir Rossi; Fernandes, João Batista; Vieira, Paulo Cezar; Vendramin, José Djair; Machado, Marcos Antônio

    2016-06-22

    The chemical composition of volatile oils from 22 genotypes of Citrus and related genera was poorly differentiated, but chemometric techniques have clarified the relationships between the 22 genotypes, and allowed us to understand their resistance to D. citri. The most convincing similarities include the synthesis of (Z)-β-ocimene and (E)-caryophyllene for all 11 genotypes of group A. Genotypes of group B are not uniformly characterized by essential oil compounds. When stimulated with odor sources of 22 genotypes in a Y-tube olfactometer D. citri preferentially entered the arm containing the volatile oils of Murraya paniculata, confirming orange jasmine as its best host. C. reticulata × C. sinensis was the least preferred genotype, and is characterized by the presence of phytol, (Z)-β-ocimene, and β-elemene, which were not found in the most preferred genotype. We speculate that these three compounds may act as a repellent, making these oils less attractive to D. citri.

  3. Nested-quantitative PCR approach with improved sensitivity for the detection of low titer levels of Candidatus Liberibacter asiaticus in the Asian citrus psyllid, Diaphorina citri Kuwayama.

    Science.gov (United States)

    Coy, M R; Hoffmann, M; Kingdom Gibbard, H N; Kuhns, E H; Pelz-Stelinski, K S; Stelinski, L L

    2014-07-01

    Candidatus Liberibacter asiaticus (CLas) is a phloem-limited bacterium transmitted by the Asian citrus psyllid, Diaphorina citri, and the presumptive causal agent of citrus greening disease. The current method of detection for CLas within plant and insect samples is by a presence/absence qPCR assay using the CLas 16S rDNA gene target. Although qPCR is highly sensitive, low bacterial titers or suboptimal qPCR conditions can result in false-negatives. Using a nested qPCR assay, we determined the false-negative rate of the 16S presence/absence qPCR assay was greater than 50%. Studies to determine the performance parameters of the qPCR assays for CLas 16S and Wingless (Wg), the D. citri endogenous gene, using plasmid and psyllid DNA, revealed suboptimal and variable performance of the 16S assay in psyllid samples. Average efficiencies and sensitivity limits of the plasmid assays were 99.0% and 2.7 copies of template for Wg, respectively, and 98.5% and 2.2-22.1 copies for 16S, respectively. Variability in efficiency was significantly greater in psyllid samples for both gene targets compared to the corresponding plasmid assays, and efficiencies as low as 76% were obtained for 16S. A secondary structure analysis revealed the formation of two stem-loop structures that block the forward and probe binding sites in the 16S template, which could hinder amplification. In summary, our results suggest that suboptimal qPCR efficiency is not uncommon for the 16S presence/absence qPCR assay, which combined with lowCLas titers in some samples, could contribute significantly to the under-reporting of CLas infection in psyllid and plant samples.

  4. Detection of citrus huanglongbing-associated 'Candidatus Liberibacter asiaticus' in citrus and Diaphorina citri in Pakistan, seasonal variability, and implications for disease management.

    Science.gov (United States)

    Razi, Muhammad F; Keremane, Manjunath L; Ramadugu, Chandrika; Roose, Mikeal; Khan, Iqrar A; Lee, Richard F

    2014-03-01

    We report the detection of the huanglongbing (HLB)-associated bacterium 'Candidatus Liberibacter asiaticus' from both plants and insects in Pakistan and the seasonal variability in the numbers of 'Ca. L. asiaticus'-positive psyllid vector, Diaphorina citri. Our studies showed that 'Ca. L. asiaticus' was detectable from trees in areas with maximum temperatures reaching nearly 50°C (average maximum of 42°C). However, the bacterium was present at very low levels in psyllids both in summer (June to August) and autumn (September to November) in contrast to reports from Florida, where the bacterium was detectable at very high levels during October to November. We hypothesize that hot summer temperatures in Pakistan may interfere with acquisition and replication of 'Ca. L. asiaticus' in psyllids and may lead to dead or non transmissible 'Ca. L. asiaticus' in plants. Psyllid counts were very low in both summer and winter, showed a population peak ('Ca. L. asiaticus'-positive vectors) in spring, and showed a larger peak ('Ca. L. asiaticus'-free psyllids) in autumn. Natural thermotherapy during hot summers and a low vector population during environmental extremes may have played a major role in long-term survival of the citrus industry in Pakistan. These results may be useful in developing management strategies for U.S. citrus industries in Texas and California.

  5. Essential Oil Variation from Twenty Two Genotypes of Citrus in Brazil—Chemometric Approach and Repellency Against Diaphorina citri Kuwayama

    Directory of Open Access Journals (Sweden)

    Moacir dos Santos Andrade

    2016-06-01

    Full Text Available The chemical composition of volatile oils from 22 genotypes of Citrus and related genera was poorly differentiated, but chemometric techniques have clarified the relationships between the 22 genotypes, and allowed us to understand their resistance to D. citri. The most convincing similarities include the synthesis of (Z-β-ocimene and (E-caryophyllene for all 11 genotypes of group A. Genotypes of group B are not uniformly characterized by essential oil compounds. When stimulated with odor sources of 22 genotypes in a Y-tube olfactometer D. citri preferentially entered the arm containing the volatile oils of Murraya paniculata, confirming orange jasmine as its best host. C. reticulata × C. sinensis was the least preferred genotype, and is characterized by the presence of phytol, (Z-β-ocimene, and β-elemene, which were not found in the most preferred genotype. We speculate that these three compounds may act as a repellent, making these oils less attractive to D. citri.

  6. 柑橘木虱对柑橘不同品种的趋性%Diaphorina citri's Gost Preference on Thirteen Citrus Varieties

    Institute of Scientific and Technical Information of China (English)

    陈建利; 阮传清; 刘波; 范国成; 段永平; D.HALL

    2011-01-01

    用2种方法测定了柑橘木虱Diaphorina citri Kuw.成虫对13个柑橘品种离体嫩梢的选择性.结果表明,对不同品种进行两两组合的选择性试验,柑橘木虱停靠在处理组合早金和四季斑叶橘的虫量占总虫量的23%和1%,停靠在处理组合台湾玫瑰橙和纽荷尔脐橙上的虫量分别占总虫量的33%和20%,停靠在处理组合日辉和佩奇甜橙上虫量分别占总虫量的38%和15%,两两之间差异显著.将13个品种组合在一起进行选择性试验,结果表明柑橘木虱停靠在纽荷尔、福橘和佩奇甜橙上的成虫数量为每梢3.2~6.6只·d-1,显著高于木虱在其余10个品种枝梢上的停靠数量.福橘是柑橘黄龙病高发品种的原因是其对柑橘木虱的吸引力较强.%Under laboratory conditions, preference of adult Asian Citrus Psyllid (ACP), Diaphorina citri Kuw, for infestation among 13 citrus varieties was determined with two experimentations. Experiment A applied every 2 detached shoots of different citrus varieties and put together for choice. Experiment B used the detached shoots of all tested citrus varieties and put together for choice. The results of Experiment A revealed that the percentage of ACP adults on each shoot of Citrus sinensis (L.) Osbeck cv. Earlygold and C. medurensis (C. mitis) cv. Varigated Calamondin were 23 % and 1 %, respectively, which was significantly different from each other. The percentage on C. sinensis Osbeck cv. Taiwanmeiguicheng (33%) and cv. Newhall Navel Orange (20%) were also significantly different from each other. Significant different percentages of ACP adults were also observed between C. reticulata C. paradisi cv. Rihui (38%) and C. reticulata C. grandis cv. Page (15%). In Experiment B, the number of ACP on the shoots of C. sinensis Osbeck cv. Newhall Navel Orange, C. tangerina Tanaka cv. Fuju and C. reticulata Citrusgrandis cv. Page was 3.2-6.6 adults · shoot-day-1 , which were significantly higher than that

  7. Advances in Biological Control of Citrus Psyllid Diaphorina citri, a Vector Insect of Citrus Huanglongbing Disease%黄龙病媒介昆虫柑橘木虱生物防治新进展

    Institute of Scientific and Technical Information of China (English)

    代晓彦; 任素丽; 周雅婷; 任顺祥; 邱宝利

    2014-01-01

    The citrus psyllid, Diaphorina citri Kuwayama, is the primary vector of citrus Huanglongbing disease, Candidatus Liberibacter asiaticus, which has the huge potential threats to the development of citrus industry in China. Based on the comprehensive understanding of D. citri control technologies, we reviewed the latest advances in biological control of D. citri and forecasted its future prospects. The main biological control resources of citrus psyllid currently are natural enemy insects and entomopathogenic fungi, among which predatory ladybeetles, lacewings, spiders and hover flies are the dominant species of predator, while Tamarixia radiata and Diaphorencyrtus aligarhensis are the two dominant parasitoids in China. Several representative entomopathogenic fungi include Lecanicillium lecanii, Isaria fumosorosea, Beauveria bassiana and Hirsutella citriformis have been reported with high virulence to citrus psyllid. This current review is expected to provide technical support and theoretical reference for efficient and sustainable control of citrus psyllid in the future.%柑橘木虱Diaphorina citri Kuwayama是柑橘黄龙病的重要媒介昆虫,对我国柑橘产业的健康发展潜在着巨大的威胁。本文在综合国内柑橘木虱防控技术的基础上,对近年来柑橘木虱生物防治的最新研究进展进行了综述,并且对其生物防治的发展前景给予了展望。目前应用于柑橘木虱生物防治的天敌资源主要是天敌昆虫及昆虫病原真菌,其中捕食性天敌主要包括瓢虫、食蚜蝇、草蛉和蜘蛛,寄生性天敌昆虫主要包括啮小蜂和跳小峰;昆虫病原真菌主要包括蜡蚧轮枝菌、拟青霉、白僵菌、桔形被毛孢等。期望通过对柑橘木虱生物防治工作的综述,为我国今后柑橘木虱的科学、高效防控及其可持续治理提供参考,不断推动我国柑橘产业的健康发展。

  8. A “walker” tool to place Diaphorina citri (Hemiptera: Liviidae) adults at predetermined sites for bioassays of behavior in citrus (Sapindales: Rutacease) trees

    Science.gov (United States)

    A walker tool was developed to assist placement of D. citri on citrus host trees in behavioral bioassays. The walker performs better than a commonly used paintbrush tool in the proportion of successful placements and in the reduction of jumps away from the citrus leaf, although it takes about two mi...

  9. Effects of soil-applied imidacloprid on Asian citrus psyllid (Hemiptera: Psyllidae) feeding behavior

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri Kuwayama is one of the most important pests of citrus due to its status as a vector of Candidatus Liberibacter asiaticus, the bacterium associated with huanglongbing (HLB) disease. The use of insecticides for vector control is the primary method of managing...

  10. New excised-leaf assay method to test inoculativity of Asian citrus psyllid (Hemiptera: Psyllidae) with Candidatus Liberibacter asiaticus associated with citrus huanglongbing disease.

    Science.gov (United States)

    Ammar, El-Desouky; Walter, Abigail J; Hall, David G

    2013-02-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the primary vector of Candidatus Liberibacter asiaticus (Las) associated with huanglongbing, or citrus greening, the most devastating citrus (Citrus spp.) disease worldwide. Here, we developed a new "excised-leaf assay" that can speed up Las-inoculativity tests on Asian citrus psyllid from the current 3-12 mo (when using whole citrus seedlings for inoculation) to only 2-3 wk. Young adults of Asian citrus psyllid that had been reared on Las-infected plants were caged on excised healthy sweet orange [Citrus sinensis (L.) Osbeck] leaves for a 1-2-wk inoculation access periods (IAP), and then both psyllids and leaves were tested later by quantitative polymerase chain reaction (PCR). When single adults were tested per leaf, percentages of Las-positive leaves averaged 2-6% by using HLBaspr primers and 10-20% by using the more sensitive LJ900 primers. Higher proportions of Las-positive leaves were obtained with 1) higher densities of inoculating psyllids (5-10 adults per leaf), 2) longer IAPs, and 3) incubation of leaves for 1 wk postinoculation before PCR. Logistic regression analysis indicated a positive correlation between Las titer in Asian citrus psyllid adults tested singly and the probability of detecting Las in the inoculated leaves, correlations that can be very useful in epidemiological studies. Comparison between excised leaves and whole seedlings, inoculated consecutively for 1 wk each by one or a group of psyllids, indicated no significant difference between Las detection in excised leaves or whole plants. This new excised-leaf assay method saves considerable time, materials, and greenhouse space, and it may enhance vector relation and epidemiological studies on Las and potentially other Liberibacter spp. associated with huanglongbing disease.

  11. 柑橘木虱两地理种群的内共生菌群落组成及传菌能力%The endosymbiotic microbiota of two geographical populations of the Asian citrus psyllid,Diaphorina citri,in China and its ability of pathogen transmission

    Institute of Scientific and Technical Information of China (English)

    马晓芳; 张学潮; 徐海君

    2012-01-01

    柑橘黄龙病(Huanglongbing,HLB)是经柑橘木虱Diaphorina citri传播的最主要柑橘病害之一,危害严重时能对柑橘产业造成毁灭性的破坏.为了鉴定福建和海南2个地理种群柑橘木虱的内共生菌群落组成,本研究对16SrRNA部分保守序列进行PCR扩增,并利用特异性引物对不同内共生菌进行了感染率检测;另外,还通过人工接虫的方法,探索柑橘木虱成虫在带黄龙病菌蕉柑Citrus reticulata cv.Tankan 上的获菌能力,以及带菌柑橘木虱成虫对黄岩蜜橘C.reticulata cv.Subcompress的传菌能力.研究发现,这2个地理种群的柑橘木虱含有相同的内共生菌组成,包括α-Proteobacteria,Wolbachia spp.,γ-Proteobacteria,mycetocyte symbionts,β-Proteobacteria,Oxalobacter 和β-Proteobacteria,Herbaspirillum,而且这2个地理种群柑橘木虱的4种内共生菌的携带率均在95%以上.柑橘木虱成虫在带菌蕉柑上饲菌28 d后,带菌率可达到82%,而带菌柑橘木虱成虫在黄岩蜜橘上传菌75 d后,可导致橘树整体带菌.本研究为柑橘木虱的进一步研究和防虫治病途径提供了一些理论依据.%Huanglongbing (HLB) is one of the most severe citrus diseases, which is usually transmitted by the Asian citrus psyllid ( Diaphorina citri), and causes great economic loss. To understand the endosyrnbiotic microbiota of the citrus psyllid from Hainan and Fujian provinces in China, the partial conserved region of 16S rRNA was amplified by PCR, and subsequently the infection rate of each endosymbiont from two geographical populations of the citrus psyllid was detected by PCR using specific primer pairs in this stduy. Moreover, we detected the acquisition efficiency of the pathogen by adult citrus psyllid reared on Citrus reticulata cv. Tank an infected with Candidatus Liberibacter as iatic us (CLas) , and the transmission efficiency to Citrus reticulata cv, Subcompress via CLas-positive psyllids. The results demonstrated that the two

  12. 联苯菊酯等6种农药对柑桔木虱成虫室内毒力测定及田间药效试验%Field Control Experiments and Toxicity of 6 Pesticides to Asian Citrus Psylla(Diaphorina citri Kuwayama)

    Institute of Scientific and Technical Information of China (English)

    邓明学; 谭有龙; 唐美欣; 陈贵峰; 唐明丽

    2009-01-01

    Citrus psylla (Diaphorina citri) has become the most important pest in Huanglongbing (HLB) areas. Up to now; the assay was yet not reported. By means of dipping, the laboratory toxicity of 6 pesticides to adult psylla was conducted out in 2008. The LC of carbosulfan, phenthoate,bifenthrin,imidacloprid, quinalphos,Piri-miphos-methyl was 4.1mg/kg, 44.3 mg/kg, 44.3 mg/kg, 60.1 mg/kg, 73.5 mg/kg, 192.8 mg/kg, respectively. Af-ter treating of 24 h, 3d and 7 d, the adult psyUid field control efficiency of bifenthrin plus azone 12.5 mg (a.i)/kg was all 100%. the adult psyllid control efficiency of phenthoate 333 mg(a.i)/kg,quinalphos 250 mg(a.i)/kg, carbo-suffan plus azone 200(a.i)/kg was all over than 95%, while that of pirimiphos-methyl 500 mg(a.i)/kg less 80%. The adult psyllid field control efficiency of carbosulfan plus azone 200(a.i)/kg was better than that of carbosul-fan 250(a.i)/kg.%为了筛选到更多防治柑桔木虱成虫的有效药剂,于2008年采用点滴法测定了6种农药对其成虫的室内毒力,活性最高的是丁硫克百威(LC50=4.1 mg/kg),其次分别是稻丰散(LC50=44.3 mg/kg)、联苯菊酯(LC50=44.3 mg/kg)、吡虫啉(LC50=60.1 mg/kg)和喹硫磷(LC50=73.5mg/kg),最差的是甲基嘧啶磷(LC50=192.8mg/kg).田间药效试验结果表明,增效联苯菊酯EC 25.00,16.67,12.50 mg(a.i)/kg药后24 h,3,7天的防效达100%;喹硫磷EC 250 mg(a.i)/kg、稻丰散EC 500 mg(a.i)/kg、333 mg(a.i)/kg、增效丁硫克百威EW200 mg(a.i)/kg药后24 h的防效>95%;甲基嘧啶磷500 mg(a.i)/kg的防效低于80%.增效丁硫克百威EW200mg(a.i)/kg的防效高于丁硫克百威EC 250mg(a.i)/kg的防效.

  13. Host plant affects morphometric variation of Diaphorina citri (Hemiptera: Liviidae)

    Science.gov (United States)

    The Asian citrus psyllid (ACP), due to its potential to vector the pathogen causing citrus greening disease or huanglongbing, is one of the most serious citrus pests worldwide. While optimal plant cultivars for ACP oviposition and development have been determined, little is known of the influence of...

  14. Attraction of Diaphorina citri (Hemiptera: Liviidae) to colored light

    Science.gov (United States)

    The most devasting disease of citrus, huanglongbing (HLB), is spread by the insect vector the Asian Citrus Psyllid (ACP). Although ACP are known to respond to visual cues, previous studies have not documented the specific wavelengths of light attractive to ACP. The objectives of this study were to...

  15. Retracted stylets in nymphs of the Asian citrus psyllid are held externally against the clypeus by a special paired organ not found in the adults

    Science.gov (United States)

    Differences in ultrastructure of the mouthparts in nymphs and adults of the Asian citrus psyllid Diaphorina citri (Hemiptera, Psyllidae), vector of the bacterium associated with citrus huanglongbing disease, were studied using scanning electron microscopy. The number of sensilla on the labial tip in...

  16. Monitoring for Insecticide Resistance in Asian Citrus Psyllid (Hemiptera: Psyllidae) Populations in Florida.

    Science.gov (United States)

    Kanga, Lambert H B; Eason, Julius; Haseeb, Muhammad; Qureshi, Jawwad; Stansly, Philip

    2016-04-01

    The development of insecticide resistance in Asian citrus psyllid, Diaphorina citri Kuwayama, populations is a serious threat to the citrus industry. As a contribution to a resistance management strategy, we developed a glass vial technique to monitor field populations of Asian citrus psyllid for insecticide resistance. Diagnostic concentrations needed to separate susceptible genotypes from resistant individuals were determined for cypermethrin (0.5 μg per vial), malathion (1.0 μg per vial), diazinon (1.0 μg per vial), carbaryl (1.0 μg per vial), carbofuran (0.1 μg per vial), methomyl (1.0 μg per vial), propoxur (1.0 μg per vial), endosulfan (1.0 μg per vial), imidacloprid (0.5 μg per vial), acetamiprid (5.0 μg per vial), chlorfenapyr (2.5 μg per vial), and fenpyroximate (2.5 μg per vial). In 2014, resistance to two carbamate insecticides (carbaryl and carbofuran), one organophosphate (malathion), one pyrethroid (cypermethrin), and one pyrazole (fenpyroximate) was detected in field populations of Asian citrus psyllid in Immokalee, FL. There was no resistance detected to diazinon, methomyl, propoxur, endosulfan, imidacloprid, and chlorfenapyr. The levels of insecticide resistance were variable and unstable, suggesting that resistance could be successfully managed. The results validate the use of the glass vial bioassay to monitor for resistance in Asian citrus psyllid populations and provide the basis for the development of a resistance management strategy designed to extend the efficacy of all classes of insecticides used for control of the Asian citrus psyllid.

  17. Sampling Methods for Detection and Monitoring of the Asian Citrus Psyllid (Hemiptera: Psyllidae).

    Science.gov (United States)

    Monzo, C; Arevalo, H A; Jones, M M; Vanaclocha, P; Croxton, S D; Qureshi, J A; Stansly, P A

    2015-06-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is a key pest of citrus due to its role as vector of citrus greening disease or "huanglongbing." ACP monitoring is considered an indispensable tool for management of vector and disease. In the present study, datasets collected between 2009 and 2013 from 245 citrus blocks were used to evaluate precision, sensitivity for detection, and efficiency of five sampling methods. The number of samples needed to reach a 0.25 standard error-mean ratio was estimated using Taylor's power law and used to compare precision among sampling methods. Comparison of detection sensitivity and time expenditure (cost) between stem-tap and other sampling methodologies conducted consecutively at the same location were also assessed. Stem-tap sampling was the most efficient sampling method when ACP densities were moderate to high and served as the basis for comparison with all other methods. Protocols that grouped trees near randomly selected locations across the block were more efficient than sampling trees at random across the block. Sweep net sampling was similar to stem-taps in number of captures per sampled unit, but less precise at any ACP density. Yellow sticky traps were 14 times more sensitive than stem-taps but much more time consuming and thus less efficient except at very low population densities. Visual sampling was efficient for detecting and monitoring ACP at low densities. Suction sampling was time consuming and taxing but the most sensitive of all methods for detection of sparse populations. This information can be used to optimize ACP monitoring efforts.

  18. Global genetic variation in the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae) and the endosymbiont Wolbachia: links between Iran and the USA detected.

    Science.gov (United States)

    Lashkari, Mohammadreza; Manzari, Shahab; Sahragard, Ahad; Malagnini, Valeria; Boykin, Laura M; Hosseini, Reza

    2014-07-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is one of the most serious pests of citrus in the world, because it transmits the pathogen that causes citrus greening disease. To determine genetic variation among geographic populations of D. citri, microsatellite markers, mitochondrial gene cytochrome oxidase I (mtCOI) and the Wolbachia-Diaphorina, wDi, gene wsp sequence data were used to characterize Iranian and Pakistani populations. Also, a Bayesian phylogenetic technique was utilized to elucidate the relationships among the sequences data in this study and all mtCOI and wsp sequence data available in GenBank and the Wolbachia database. Microsatellite markers revealed significant genetic differentiation among Iranian populations, as well as between Iranian and Pakistani populations (FST  = 0.0428, p Mexico, Florida and Texas (USA) are similar. Wolbachia, wDi, wsp sequences were similar among Iranian populations, but different between Iranian and Pakistani populations. The South West Asia (SWA) group is the most likely source of the introduced Iranian populations of D. citri. This assertion is also supported by the sequence similarity of the Wolbachia, wDi, strains from the Florida, USA and Iranian D. citri. These results should be considered when looking for biological controls in either country. © 2013 Society of Chemical Industry.

  19. H NMR analyses of Citrus macrophylla subjected to Asian citrus psyllid (Diaphorina citri Kuwayama) feeding

    Science.gov (United States)

    The Asian citrus psyllid (ACP) is a phloem feeding insect that can host and transmit the bacterium Candidatus Liberibacter asiaticus (CLas), which is the putative causative agent of the economically important citrus disease, Huanglongbing (HLB). ACP are widespread in Florida, and are spreading in Ca...

  20. Proposal to conserve the name Phomopsis citri H.S. Fawc. (Diaporthe citri), with a conserved type, against Phomopsis citri (Sacc.) Traverso & Spessa (Ascomycota, Diaporthales, Diaporthaceae)

    Science.gov (United States)

    The name Diaporthe citri applies to a fungus that causes a disease on Citrus known as melanose or stem end rot of mature fruit after harvest and occurs widely in North America and Asia. Initially described as the illegitimate Phomopsis citri H.S. Fawc. 1912, non P. citri (Sacc.) Traverso & Spessa 19...

  1. Foliar application of biofilm formation-inhibiting compounds enhances control of citrus canker caused by Xanthomonas citri subsp. citri.

    Science.gov (United States)

    Li, Jinyun; Wang, Nian

    2014-02-01

    Citrus canker caused by the bacterium Xanthomonas citri subsp. citri is an economically important disease of citrus worldwide. Biofilm formation plays an important role in early infection of X. citri subsp. citri on host leaves. In this study, we assessed the hypothesis that small molecules inhibiting biofilm formation reduce X. citri subsp. citri infection and enhance the control of citrus canker disease. D-leucine and 3-indolylacetonitrile (IAN) were found to prevent biofilm formation by X. citri subsp. citri on different abiotic surfaces and host leaves at a concentration lower than the minimum inhibitory concentration (MIC). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis indicated that IAN repressed expression of chemotaxis/motility-related genes in X. citri subsp. citri. In laboratory experiments, planktonic and biofilm cells of X. citri subsp. citri treated with D-leucine and IAN, either alone or in combination, were more susceptible to copper (CuSO4) than those untreated. In greenhouse assays, D-leucine and IAN applied alone or combined with copper reduced both the number of canker lesions and bacterial populations of X. citri subsp. citri on citrus host leaves. This study provides the basis for the use of foliar-applied biofilm inhibitors for the control of citrus canker alone or combined with copper-based bactericides.

  2. Fortunella margarita Transcriptional Reprogramming Triggered by Xanthomonas citri subsp. citri

    Directory of Open Access Journals (Sweden)

    Khalaf Abeer A

    2011-11-01

    Full Text Available Abstract Background Citrus canker disease caused by the bacterial pathogen Xanthomonas citri subsp. citri (Xcc has become endemic in areas where high temperature, rain, humidity, and windy conditions provide a favourable environment for the dissemination of the bacterium. Xcc is pathogenic on many commercial citrus varieties but appears to elicit an incompatible reaction on the citrus relative Fortunella margarita Swing (kumquat, in the form of a very distinct delayed necrotic response. We have developed subtractive libraries enriched in sequences expressed in kumquat leaves during both early and late stages of the disease. The isolated differentially expressed transcripts were subsequently sequenced. Our results demonstrate how the use of microarray expression profiling can help assign roles to previously uncharacterized genes and elucidate plant pathogenesis-response related mechanisms. This can be considered to be a case study in a citrus relative where high throughput technologies were utilized to understand defence mechanisms in Fortunella and citrus at the molecular level. Results cDNAs from sequenced kumquat libraries (ESTs made from subtracted RNA populations, healthy vs. infected, were used to make this microarray. Of 2054 selected genes on a customized array, 317 were differentially expressed (P Conclusion Functional categorization of kumquat Xcc-responsive genes revealed an enhanced defence-related metabolism as well as a number of resistant response-specific genes in the kumquat transcriptome in response to Xcc inoculation. Gene expression profile(s were analyzed to assemble a comprehensive and inclusive image of the molecular interaction in the kumquat/Xcc system. This was done in order to elucidate molecular mechanisms associated with the development of the hypersensitive response phenotype in kumquat leaves. These data will be used to perform comparisons among citrus species to evaluate means to enhance the host immune responses

  3. The type III protein secretion system contributes to Xanthomonas citri subsp. citri biofilm formation

    KAUST Repository

    Zimaro, Tamara

    2014-04-18

    Background: Several bacterial plant pathogens colonize their hosts through the secretion of effector proteins by a Type III protein secretion system (T3SS). The role of T3SS in bacterial pathogenesis is well established but whether this system is involved in multicellular processes, such as bacterial biofilm formation has not been elucidated. Here, the phytopathogen Xanthomonas citri subsp. citri (X. citri) was used as a model to gain further insights about the role of the T3SS in biofilm formation. Results: The capacity of biofilm formation of different X. citri T3SS mutants was compared to the wild type strain and it was observed that this secretion system was necessary for this process. Moreover, the T3SS mutants adhered proficiently to leaf surfaces but were impaired in leaf-associated growth. A proteomic study of biofilm cells showed that the lack of the T3SS causes changes in the expression of proteins involved in metabolic processes, energy generation, exopolysaccharide (EPS) production and bacterial motility as well as outer membrane proteins. Furthermore, EPS production and bacterial motility were also altered in the T3SS mutants. Conclusions: Our results indicate a novel role for T3SS in X. citri in the modulation of biofilm formation. Since this process increases X. citri virulence, this study reveals new functions of T3SS in pathogenesis. 2014 Zimaro et al.; licensee BioMed Central Ltd.

  4. Complete Genome Sequences of Six Copper-Resistant Xanthomonas citri pv.?citri Strains Causing Asiatic Citrus Canker, Obtained Using Long-Read Technology

    OpenAIRE

    Richard, Damien; Boyer, Claudine; Vernière, Christian; Canteros, B.I.; Lefeuvre, Pierre; Pruvost, Olivier

    2017-01-01

    The gammaproteobacterium Xanthomonas citri pv. citri causes Asiatic citrus canker. Pathotype A strains have a broad host range, which includes most commercial citrus species, and they cause important economic losses worldwide. Control often relies on frequent copper sprays. We present here the complete genomes of six X. citri pv. citri copper-resistant strains. (Résumé d'auteur)

  5. Complete Genome Sequences of Six Copper-Resistant Xanthomonas citri pv. citri Strains Causing Asiatic Citrus Canker, Obtained Using Long-Read Technology

    Science.gov (United States)

    Richard, Damien; Boyer, Claudine; Vernière, Christian; Canteros, Blanca I.; Lefeuvre, Pierre

    2017-01-01

    ABSTRACT The gammaproteobacterium Xanthomonas citri pv. citri causes Asiatic citrus canker. Pathotype A strains have a broad host range, which includes most commercial citrus species, and they cause important economic losses worldwide. Control often relies on frequent copper sprays. We present here the complete genomes of six X. citri pv. citri copper-resistant strains. PMID:28336584

  6. Biology and management of Asian citrus psyllid, vector of the huanglongbing pathogens.

    Science.gov (United States)

    Grafton-Cardwell, Elizabeth E; Stelinski, Lukasz L; Stansly, Philip A

    2013-01-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the most important pest of citrus worldwide because it serves as a vector of "Candidatus Liberibacter" species (Alphaproteobacteria) that cause huanglongbing (citrus greening disease). All commercially cultivated citrus is susceptible and varieties tolerant to disease expression are not yet available. Onset of disease occurs following a long latent period after inoculation, and thus the pathogen can spread widely prior to detection. Detection of the pathogen in Brazil in 2004 and Florida in 2005 catalyzed a significant increase in research on D. citri biology. Chemical control is the primary management strategy currently employed, but recently documented decreases in susceptibility of D. citri to several insecticides illustrate the need for more sustainable tools. Herein, we discuss recent advances in the understanding of D. citri biology and behavior, pathogen transmission biology, biological control, and chemical control with respect to "Candidatus Liberibacter asiaticus." Our goal is to point toward integrated and biologically relevant management of this pathosystem.

  7. The cys regulon of Xanthomonas citri

    Energy Technology Data Exchange (ETDEWEB)

    Moutran, A.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

    2012-07-01

    Full text: In Escherichia coli, genes involved in metabolic pathway of sulfate and sulfonate compounds are clustered in a cys regulon, which includes three ABC transport system (operons: sbpcysWUA; ssuABC and tauABC), thirteen genes involved in the sulfur reduction (ssuDE; tauD and cysDNCHIJGK) and two regulatory proteins that belong to LysR transcription family: CysB and Cbl. Notably, a search and comparative analysis of these genes in the genomes of the citrus pathogen Xanthomonas citri and other phylogenetically related Xanthomonas species revealed the presence of genes involved with alkanesulfonate, sulfate ester and taurine, only in X. citri, suggesting that proteins from this regulon might be associated with pathogenicity in citrus. Using the molecular modeling associated with a system biology view, we modeled all the protein structures of the X. citri cys regulon as well as characterized the important residues forming the putative active sites. Comparison with orthologs from different microorganisms was made in order to get a phylogenetic relationships. We showed that proteins that are responsible for the affinity and specificity of the alkanesulfonate, sulfate and taurine transport systems conserved the residues involved in the sulfate coordination but are organized in different branches in evolution. Inside these phylogenetic branches, proteins involved in the sulfate transporter are highly conserved when compared to the others. Moreover, we identified that the taurine-binding protein (TauA) of the X. citri belongs to a different evolutionary branch from that one that described for E. coli. These differences were also noticed for components of the tau operon, including a putative new regulator. The function and mechanism of action of each protein is discussed in order to bring light for the sulfur assimilation processes and their importance for X. citri physiology. (author)

  8. Characterization of a variant of Xanthomonas citri subsp. citri that triggers a host-specific defense response.

    Science.gov (United States)

    Chiesa, María A; Siciliano, María F; Ornella, Leonardo; Roeschlin, Roxana A; Favaro, María A; Delgado, Natalia Pino; Sendín, Lorena N; Orce, Ingrid G; Ploper, L Daniel; Vojnov, Adrian A; Vacas, José Gadea; Filippone, María P; Castagnaro, Atilio P; Marano, María R

    2013-06-01

    Citrus is an economically important fruit crop that is severely afflicted by Asiatic citrus bacterial canker (CBC), a disease caused by the phytopathogen Xanthomonas citri subsp. citri (X. citri). To gain insight into the molecular epidemiology of CBC, 42 Xanthomonas isolates were collected from a range of Citrus spp. across 17 different orchards in Tucumán, Argentina and subjected to molecular, biochemical, and pathogenicity tests. Analysis of genome-specific X. citri markers and DNA polymorphisms based on repetitive elements-based polymerase chain reaction showed that all 42 isolates belonged to X. citri. Interestingly, pathogenicity tests showed that one isolate, which shares >90% genetic similarity to the reference strain X. citri T, has host range specificity. This new variant of X. citri subsp. citri, named X. citri A(T), which is deficient in xanthan production, induces an atypical, noncankerous chlorotic phenotype in Citrus limon and C. paradisi and weak cankerous lesions in C. aurantifolia and C. clementina leaves. In C. limon, suppression of canker development is concomitant with an oxidative burst; xanthan is not implicated in the phenotype induced by this interaction, suggesting that other bacterial factors would be involved in triggering the defense response.

  9. Evaluation of systemic neonicotinoid insecticides for the management of the Asian citrus psyllid Diaphorina citri on containerized citrus.

    Science.gov (United States)

    Byrne, Frank J; Daugherty, Matthew P; Grafton-Cardwell, Elizabeth E; Bethke, James A; Morse, Joseph G

    2017-03-01

    Studies were conducted to evaluate uptake and retention of three systemic neonicotinoid insecticides, dinotefuran, imidacloprid and thiamethoxam, in potted citrus nursery plants treated at standard label rates. Infestation of these plants placed at a field site with moderate levels of Asian citrus psyllid (ACP) was monitored for 14 weeks following treatments, and insecticide residues in leaf tissue were quantified using enzyme-linked immunosorbent assay (ELISA). Bioassays were conducted using leaves harvested on various dates post-treatment to compare the efficacies of residues against adult ACP. Residues of the three neonicotinoids were detected in leaf tissues within 1 week after treatment. Peak concentrations established at 1 week for imidacloprid and dinotefuran and at 2 weeks for thiamethoxam. Imidacloprid and thiamethoxam outperformed the control and dinotefuran treatments at protecting trees from infestations by ACP eggs and nymphs. For a given insecticide concentration in leaf tissue, thiamethoxam induced the highest mortality of the three insecticides, and dinotefuran was the least toxic. If the time needed to achieve effective thresholds of a systemic neonicotinoid is known, treatments at production facilities could be scheduled that would minimize unnecessary post-treatment holding periods and ensure maximum retention of effective concentrations after the plants have shipped to retail outlets. The rapid uptake of the insecticides and retention at effective concentrations in containerized citrus suggest that the current 30 day post-treatment shipping restriction from production facilities to retail outlets outside of quarantine could be shortened to 14 days. Thiamethoxam should be added to the list of approved nursery treatments. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  10. Asian citrus psyllids and shade: Survival of Diaphorina citri on Murraya exotica foliage exposed to different levels of light

    Science.gov (United States)

    Using shade cloths of different densities, potted individuals of Murraya exotica (Rutaceae) were exposed to levels of photosynthetic radiation (PAR) varying from 48 µmol photons per meter2 per second to 1562 µmol photons per meter2 per second (average of three readings taken at noon during condition...

  11. Silencing abnormal wing disc gene of the Asian citrus psyllid, Diaphorina citri disrupts adult wing development and increases nymph mortality.

    Directory of Open Access Journals (Sweden)

    Ibrahim El-Shesheny

    Full Text Available Huanglongbing (HLB causes considerable economic losses to citrus industries worldwide. Its management depends on controlling of the Asian citrus Psyllid (ACP, the vector of the bacterium, Candidatus Liberibacter asiaticus (CLas, the causal agent of HLB. Silencing genes by RNA interference (RNAi is a promising tool to explore gene functions as well as control pests. In the current study, abnormal wing disc (awd gene associated with wing development in insects is used to interfere with the flight of psyllids. Our study showed that transcription of awd is development-dependent and the highest level was found in the last instar (5(th of the nymphal stage. Micro-application (topical application of dsRNA to 5(th instar of nymphs caused significant nymphal mortality and adult wing-malformation. These adverse effects in ACP were positively correlated with the amounts of dsRNA used. A qRT-PCR analysis confirmed the dsRNA-mediated transcriptional down-regulation of the awd gene. Significant down-regulation was required to induce a wing-malformed phenotype. No effect was found when dsRNA-gfp was used, indicating the specific effect of dsRNA-awd. Our findings suggest a role for awd in ACP wing development and metamorphosis. awd could serve as a potential target for insect management either via direct application of dsRNA or by producing transgenic plants expressing dsRNA-awd. These strategies will help to mitigate HLB by controlling ACP.

  12. Comparison of DNA extraction methods for detection of citrus huanglongbing in Colombia

    Directory of Open Access Journals (Sweden)

    Jorge Evelio Ángel

    2014-04-01

    Full Text Available Four DNA citrus plant tissue extraction protocols and three methods of DNA extraction from vector psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae were compared as part of the validation process and standardization for detection of huanglongbing (HLB. The comparison was done using several criterias such as integrity, purity and concentration. The best quality parameters presented in terms of extraction of DNA from plant midribs tissue of citrus, were cited by Murray and Thompson (1980 and Rodríguez et al. (2010, while for the DNA extraction from psyllid vectors of HLB, the best extraction method was suggested by Manjunath et al.(2008.

  13. Scientific Opinion on the risk to plant health of Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii for the EU territory

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Plant Health (PLH

    2014-02-01

    Full Text Available The Panel conducted a pest risk assessment for Xanthomonas campestris (all strains pathogenic to Citrus for the EU territory and an assessment of the effectiveness of present EU requirements against Xanthomonas strains pathogenic to citrus. The risk assessment was conducted under the scenario of absence of the current specific EU plant health legislation and the assumption that citrus-exporting countries apply measures to reduce yield and quality losses. Risk reduction options were systematically identified and evaluated. The strains of X. campestris pathogenic to citrus have been reclassified as four distinct infraspecific taxa within two species: X. citri and X. alfalfae. Only two pathovars (X. citri pv. citri and X. citri pv. aurantifolii are responsible for the citrus bacterial canker thatpresents a major risk for the citrus industry in the EU. Seven entry pathways have been identified and evaluated. The likelihood of entry was rated unlikely for fruit, very likely for fruit plants for planting, moderately likely for ornamental plants for planting and unlikely for leaves and twigs. The uncertainty of probability of entry was rated as high. The probability of establishment was rated as moderately likely to likely with a medium uncertainty because host plants are widely present in EU areas where environmental conditions are suitable. Once established, spread would be likely with a low uncertainty. The impact of the disease, even if control measures are applied, was rated as moderate to major with a medium uncertainty. The disease would cause yield losses in areas where citrus is the main crop, increase the need for control measures and create environmental problems.The combined EU regulations have been shown to be effective in preventing the introduction of X. citri pv. citri or X. citri pv. aurantifolii in the EU, as no outbreaks of citrus canker in the EU territory have been reported.

  14. New corological and biological data of the Red Gum Lerp Psyllid, Glycaspis brimblecombei Moore, 1964 in Italy (Hemiptera, Psyllidae

    Directory of Open Access Journals (Sweden)

    Ricardo Jiménez-Peydró

    2011-03-01

    Full Text Available Glycaspis brimblecombei Moore, 1964 is a psyllid (Hemiptera: Psyllidae pest of Eucalyptus, native toAustralia and first recorded in Europe: Spain in 2008 and more recently (2010 in Italy. The present paper dealswith recent research, carried out in central Italy, with new data on the distribution and biology of this species.

  15. Selection of Small Synthetic Antimicrobial Peptides Inhibiting Xanthomonas citri subsp. citri Causing Citrus Canker

    Science.gov (United States)

    Choi, Jeahyuk; Park, Euiho; Lee, Se-Weon; Hyun, Jae-Wook; Baek, Kwang-Hyun

    2017-01-01

    Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fourteen hexapeptides were selected by using positional scanning of synthetic peptide combinatorial libraries. Each hexapeptide showed different antimicrobial spectrum against Bacillus, Pseudomonas, Xanthomonas, and Candida species. Intriguingly, BHC10 showed bactericidal activity exclusively on Xanthomonas citri subsp. citri (Xcc), while BHC7 was none-active exclusively against two Pseudomonas spp. at concentration of 100 μg/ml suggesting potential selectivity constrained in hexapeptide frame. Three hexapeptides, BHC02, 06 and 11, showed bactericidal activities against various Xcc strains at concentration of 10 μg/ml. When they were co-infiltrated with pathogens into citrus leaves the disease progress was suppressed significantly. Further study would be needed to confirm the actual disease control capacity of the selected hexapeptides. PMID:28167892

  16. Glycaspis brimblecombei (Hem.: Psyllidae, the invasive red gum lerp psyllid recorded in Italy

    Directory of Open Access Journals (Sweden)

    Garonna AP

    2011-05-01

    Full Text Available During 2010 in central and southern Italy (Campania, Basilicata and Latium, the invasive red gum lerp psyllid, Glycaspis brimblecombei Moore (Hemiptera: Psyllidae, has been recorder for the first time in Italy. The psyllid was detected on Eucalyptus camaldulensis in several sampling sites. The host plant, like other species of the genus Eucalyptus, is very common in Italy as ornamental and forest species. The native country of G. brimblecombei is Australia, from which the psyllid spread throughout the world in the last decade. In the Palaearctic Region the species was first collected from the Iberian Peninsula in 2007. In this note we report some observations on the biology and distribution of this species in the new colonized Italian areas. Generalist predators, such as wasps and pirate bugs, were observed attacking G. brimblecombei, however no parasitoid activity has been detected.

  17. Leaf Damaged by Nymph of Cardiaspina albitextura and Cardiaspina retator (Hemiptera: Psyllidae

    Directory of Open Access Journals (Sweden)

    NISMAH

    2008-06-01

    Full Text Available Cardispina (Hemiptera: Pshyllidae is one of the most important genera of the Australian Psyllidae, owing to the spectacular damage to Eucalyptus spp. The increase of C. albitextura and C. retator populations in high numbers caused severe damage on leaves of Eucalyptus camaldulensis. Both species have similarities, i.e. they occur on the same host plant and prefer mature leaves of E. camaldulensis for oviposition and feeding sites. They might thus be more likely to show competition than the less closely related taxa. The purpose of the study was to examine degree of the resource use by measuring the area of leaves damaged by the nymphs of C. albitextura and C. retator. The results indicated that the nymph of C. retator caused damage twice more damages than that of C. albitextura. The leaves area damaged by the females of both species was greater than that by the males.

  18. Coccidoxenoides perminutus parasitizing Planococcus citri on vine in Brazil

    Directory of Open Access Journals (Sweden)

    Maria Herlândia de Araújo Fernandes

    2016-07-01

    Full Text Available ABSTRACT: From those of the mealybug family, the citrus mealybug, Planococcus citriRisso, 1813 (Hemiptera: Pseudococcidae is frequently held responsible for various diseases occurring in vineyards. This is a first time report in Brazil, on the occurrence of a parasitoid controlling P. citri. In our search for the natural enemies of P. citri, bunches of grapes with the vine were collected every week for one month, in those regions where table grapes were produced, like Petrolina, Pernambuco, Brazil. In these samples, among the natural pest infestations Coccidoxenoides perminutusGirault, 1915 (Hymenoptera: Encyrtidae was the only natural enemy species reported. In this research, we attempted to study the potential of parasitism as well as the strategies that this natural agent uses in pest control.

  19. Ultraviolet irradiation mutagenesis and recombination in spiroplasma citri

    Energy Technology Data Exchange (ETDEWEB)

    Labarere, J.; Barroso, G. (Bordeaux-2 Univ., 33 (France))

    1984-09-01

    A method for obtaining and screening uv-induced mutants from Spiroplasma citri is described. Lethality response curves showed that S. citri is more sensitive to uv irradiation than are other microorganisms. The presence of a shoulder in the lethality response curve showed the existence of systems able to repair uv-induced DNA damages. Toxic-resistant mutants have been obtained. A uv fluence equal to 10 J/m/sup 2/ multiplied by 2.5x10/sup 3/ gave the spontaneous mutation frequency. Arsenic acid- and xylitol-resistant mutants were used to investigate transfer of genetic information in S. citri. After 90 min of incubation, the recombination frequency was 5x10/sup -5/.

  20. treA Codifies for a Trehalase with Involvement in Xanthomonas citri subsp. citri Pathogenicity

    Science.gov (United States)

    Alexandrino, André Vessoni; Goto, Leandro Seiji; Novo-Mansur, Maria Teresa Marques

    2016-01-01

    Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (Xcc), is a severe disease of citrus. Xcc presents broad spectrum of citrus hosts including economically important species whereas X. fuscans subsp. aurantifolii–type C (XauC) causes a milder disease and only infects Citrus aurantifolia. Trehalase catalyzes hydrolysis of the disaccharide trehalose, a sugar that has been reported to be related to Xcc pathogenicity. We expressed the recombinant gene product and assessed Xcc trehalase structural and kinetics data. The recombinant protein presented 42.7% of secondary structures in α-helix and 13% in β-sheets, no quaternary structure in solution, and Michaelis-Menten constant (KM) of 0.077 mM and Vmax 55.308 μMol glucose.min-1.mg protein-1 for trehalose. A Xcc mutant strain (XccΔtreA) was produced by gene deletion from Xcc genome. Enzymatic activity of trehalase was determined in Xcc, XauC and XccΔtreA cellular lysates, showing the highest values for XauC in in vitro infective condition and no activity for XccΔtreA. Finally, leaves of Citrus aurantifolia infected with XccΔtreA showed much more drenching and necrosis than those infected by wild type Xcc. We concluded that trehalase contributes to alleviate bacterial virulence and that inability for trehalose hydrolysis may promote higher Xcc infectivity. PMID:27611974

  1. treA Codifies for a Trehalase with Involvement in Xanthomonas citri subsp. citri Pathogenicity.

    Science.gov (United States)

    Alexandrino, André Vessoni; Goto, Leandro Seiji; Novo-Mansur, Maria Teresa Marques

    2016-01-01

    Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (Xcc), is a severe disease of citrus. Xcc presents broad spectrum of citrus hosts including economically important species whereas X. fuscans subsp. aurantifolii-type C (XauC) causes a milder disease and only infects Citrus aurantifolia. Trehalase catalyzes hydrolysis of the disaccharide trehalose, a sugar that has been reported to be related to Xcc pathogenicity. We expressed the recombinant gene product and assessed Xcc trehalase structural and kinetics data. The recombinant protein presented 42.7% of secondary structures in α-helix and 13% in β-sheets, no quaternary structure in solution, and Michaelis-Menten constant (KM) of 0.077 mM and Vmax 55.308 μMol glucose.min-1.mg protein-1 for trehalose. A Xcc mutant strain (XccΔtreA) was produced by gene deletion from Xcc genome. Enzymatic activity of trehalase was determined in Xcc, XauC and XccΔtreA cellular lysates, showing the highest values for XauC in in vitro infective condition and no activity for XccΔtreA. Finally, leaves of Citrus aurantifolia infected with XccΔtreA showed much more drenching and necrosis than those infected by wild type Xcc. We concluded that trehalase contributes to alleviate bacterial virulence and that inability for trehalose hydrolysis may promote higher Xcc infectivity.

  2. A protein expression system for tandem affinity purification in Xanthomonas citri subsp. citri

    Directory of Open Access Journals (Sweden)

    Giordanni C. Dantas

    2016-06-01

    Full Text Available Abstract Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp. citri (Xac, is one of the most devastating diseases to affect citrus crops. There is no treatment for citrus canker; effective control against the spread of Xac is usually achieved by the elimination of affected plants along with that of asymptomatic neighbors. An in depth understanding of the pathogen is the keystone for understanding of the disease; to this effect we are committed to the development of strategies to ease the study of Xac. Genome sequencing and annotation of Xac revealed that ∼37% of the genome is composed of hypothetical ORFs. To start a systematic characterization of novel factors encoded by Xac, we constructed integrative-vectors for protein expression specific to this bacterium. The vectors allow for the production of TAP-tagged proteins in Xac under the regulation of the xylose promoter. In this study, we show that a TAP-expression vector, integrated into the amy locus of Xac, does not compromise its virulence. Furthermore, our results also demonstrate that the polypeptide TAP can be overproduced in Xac and purified from the soluble phase of cell extracts. Our results substantiate the use of our vectors for protein expression in Xac thus contributing a novel tool for the characterization of proteins and protein complexes generated by this bacterium in vivo.

  3. Bioactive Organocopper Compound from Pseudomonas aeruginosa Inhibits the Growth of Xanthomonas citri subsp. citri

    Science.gov (United States)

    de Oliveira, Admilton G.; Spago, Flavia R.; Simionato, Ane S.; Navarro, Miguel O. P.; da Silva, Caroline S.; Barazetti, André R.; Cely, Martha V. T.; Tischer, Cesar A.; San Martin, Juca A. B.; de Jesus Andrade, Célia G. T.; Novello, Cláudio R.; Mello, João C. P.; Andrade, Galdino

    2016-01-01

    Citrus canker is a very destructive disease of citrus species. The challenge is to find new compounds that show strong antibiotic activity and low toxicity to plants and the environment. The objectives of the present study were (1) to extract, purify and evaluate the secondary metabolites with antibiotic activity produced by Pseudomonas aeruginosa LV strain in vitro against Xanthomonas citri subsp. citri (strain 306), (2) to determine the potential of semi-purified secondary metabolites in foliar application to control citrus canker under greenhouse conditions, and (3) to identify antibiotic activity in orange leaf mesophyll infected with strain 306, by electron microscopy. Two pure bioactive compounds were isolated, an organocopper antibiotic compound (OAC) and phenazine-1-carboxamide. Phenazine-1-carboxamide did not show any antibiotic activity under the experimental conditions used in this study. The OAC showed a high level of antibiotic activity with a minimum inhibitory concentration of 0.12 μg mL-1. In greenhouse tests for control of citrus canker in orange trees, the semi-purified fraction F3d reduced lesion formation by about 97%. The concentration used was 500 times lower than that for the recommended commercial copper-based product. Electron microscopy showed that F3d altered the exopolysaccharide matrix and caused cell lysis of the pathogen inside the citrus canker lesions. These results suggest that secondary metabolites produced by inducing P. aeruginosa LV strain have a high potential to be used as a bioproduct to control citrus canker. PMID:26903992

  4. In Vitro and In Vivo Antibacterial Activity of Some Organic and Inorganic Salts Against Asiatic Citrus Canker Agent Xanthomonas Citri Subsp. Citri

    Directory of Open Access Journals (Sweden)

    Vahideh Hasabi

    2014-06-01

    Full Text Available Asiatic citrus canker caused by Xanthomonas citri subsp. citri is becoming a disease of high economic impact, affecting all types of important citrus crops. In this study, the potential antibacterial activity of ten organic and inorganic salts on X. citri subsp. citri and on citrus canker disease development was evaluated. Among the salt compounds, copper, iron and zinc inorganic salts particularly zinc (with the highest diameter of inhibition, the lowest MIC and MBC values and the highest bacterial growth inhibitory effect had direct antibacterial activity and strongly reduced the development of canker disease and bacterial population of lime plants.

  5. Insights into xanthomonas axonopodis pv. Citri biofilm through proteomics

    KAUST Repository

    Zimaro, Tamara

    2013-08-07

    Background: Xanthomonas axonopodis pv. Citri (X. a. pv. Citri) causes citrus canker that can result in defoliation and premature fruit drop with significant production losses worldwide. Biofilm formation is an important process in bacterial pathogens and several lines of evidence suggest that in X. a. pv. Citri this process is a requirement to achieve maximal virulence since it has a major role in host interactions. In this study, proteomics was used to gain further insights into the functions of biofilms. Results: In order to identify differentially expressed proteins, a comparative proteomic study using 2D difference gel electrophoresis was carried out on X. a. pv. Citri mature biofilm and planktonic cells. The biofilm proteome showed major variations in the composition of outer membrane proteins and receptor or transport proteins. Among them, several porins and TonB-dependent receptor were differentially regulated in the biofilm compared to the planktonic cells, indicating that these proteins may serve in maintaining specific membrane-associated functions including signaling and cellular homeostasis. In biofilms, UDP-glucose dehydrogenase with a major role in exopolysaccharide production and the non-fimbrial adhesin YapH involved in adherence were over-expressed, while a polynucleotide phosphorylase that was demonstrated to negatively control biofilm formation in E. coli was down-regulated. In addition, several proteins involved in protein synthesis, folding and stabilization were up-regulated in biofilms. Interestingly, some proteins related to energy production, such as ATP-synthase were down-regulated in biofilms. Moreover, a number of enzymes of the tricarboxylic acid cycle were differentially expressed. In addition, X. a. pv. Citri biofilms also showed down-regulation of several antioxidant enzymes. The respective gene expression patterns of several identified proteins in both X. a. pv. Citri mature biofilm and planktonic cells were evaluated by

  6. The Effect of Citrus Essential Oils and Their Constituents on Growth of Xanthomonas citri subsp. citri.

    Science.gov (United States)

    Mirzaei-Najafgholi, Hossein; Tarighi, Saeed; Golmohammadi, Morteza; Taheri, Parissa

    2017-04-14

    Citrus bacterial canker (CBC) caused by Xanthomonas citri subsp. citri (Xcc), is the most devastating of the citrus diseases worldwide. During our study, we found that Essential oils (EOs) of some citrus cultivars are effective on Xcc. Therefore, it prompted us to determine the plant metabolites responsible for the antibacterial properties. We obtained EOs from some locally cultivated citrus by using a Clevenger apparatus and their major constituents were identified by gas chromatography/mass spectrometry (GC-MS). The effect of Citrus aurantium, C. aurantifolia, Fortunella sp. EOs and their major constituents were evaluated against Xcc-KVXCC1 using a disk diffusion assay. Minimal inhibitory and bactericidal concentration of the EOs and their constituents were determined using the broth microdilution method. C. aurantium, C. aurantifolia Eos, and their major constituents including citral, linalool, citronellal, geraniol, α-terpineol, and linalyl acetate indicated antibacterial effects against Xcc. The C. aurantifolia EO and citral showed the highest antibacterial activity among the tested EOs and constituents with inhibition zones of 15 ± 0.33 mm and 16.67 ± 0.88 mm, respectively. Synergistic effects of the constituents were observed between α-terpineol-citral, citral-citronellal, citral-geraniol, and citronellal-geraniol by using a microdilution checkerboard assay. Transmission electron microscopy revealed that exposure of Xcc cells to citral caused cell wall damage and altered cytoplasmic density. We introduced C. aurantifolia and C. aurantium EOs, and their constituents citral, α-terpineol, citronellal, geraniol, and linalool as possible control agents for CBC.

  7. Proteome of the phytopathogen Xanthomonas citri subsp. citri: a global expression profile

    Directory of Open Access Journals (Sweden)

    Ferro Jesus A

    2010-11-01

    Full Text Available Abstract Background Citrus canker is a disease caused by Xantomonas citri subsp.citri (Xac, and has emerged as one of the major threats to the worldwide citrus crop because it affects all commercial citrus varieties, decreases the production and quality of the fruits and can spread rapidly in citrus growing areas. In this work, the first proteome of Xac was analyzed using two methodologies, two-dimensional liquid chromatography (2D LC and tandem mass spectrometry (MS/MS. Results In order to gain insight into the metabolism of Xac, cells were grown on two different media (NB - Nutrient Broth and TSE - Tryptone Sucrose broth enriched with glutamic acid, and proteins were proteolyzed with trypsin and examined by 2D LC-MS/MS. Approximately 39% of all predicted proteins by annotation of Xac were identified with their component peptides unambiguously assigned to tandem mass spectra. The proteins, about 1,100, were distributed in all annotated functional categories. Conclusions This is the first proteomic reference map for the most aggressive strain of Xanthomonas pathogen of all orange varieties. The compilation of metabolic pathways involved with bacterial growth showed that Xac expresses a complete central and intermediary metabolism, replication, transcription and translation machineries and regulation factors, distinct membrane transporters (ABC, MFS and pumps and receptors (MCP, TonB dependent and metabolites acquisition, two-component systems (sensor and regulatory components and response regulators. These data corroborate the growth curve in vitro and are the first reports indicating that many of these genome annotated genes are translated into operative in Xac. This proteomic analysis also provided information regarding the influence of culture medium on growth and protein expression of Xac.

  8. Non-host defense response in a novel Arabidopsis-Xanthomonas citri subsp. citri pathosystem.

    Directory of Open Access Journals (Sweden)

    Chuanfu An

    Full Text Available Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc, is one of the most destructive diseases of citrus. Progress of breeding citrus canker-resistant varieties is modest due to limited resistant germplasm resources and lack of candidate genes for genetic manipulation. The objective of this study is to establish a novel heterologous pathosystem between Xcc and the well-established model plant Arabidopsis thaliana for defense mechanism dissection and resistance gene identification. Our results indicate that Xcc bacteria neither grow nor decline in Arabidopsis, but induce multiple defense responses including callose deposition, reactive oxygen species and salicylic aicd (SA production, and defense gene expression, indicating that Xcc activates non-host resistance in Arabidopsis. Moreover, Xcc-induced defense gene expression is suppressed or attenuated in several well-characterized SA signaling mutants including eds1, pad4, eds5, sid2, and npr1. Interestingly, resistance to Xcc is compromised only in eds1, pad4, and eds5, but not in sid2 and npr1. However, combining sid2 and npr1 in the sid2npr1 double mutant compromises resistance to Xcc, suggesting genetic interactions likely exist between SID2 and NPR1 in the non-host resistance against Xcc in Arabidopsis. These results demonstrate that the SA signaling pathway plays a critical role in regulating non-host defense against Xcc in Arabidopsis and suggest that the SA signaling pathway genes may hold great potential for breeding citrus canker-resistant varieties through modern gene transfer technology.

  9. Amplified fragment length polymorphism and multilocus sequence analysis-based genotypic relatedness among pathogenic variants of Xanthomonas citri pv. citri and Xanthomonas campestris pv. bilvae.

    Science.gov (United States)

    Bui Thi Ngoc, Lan; Vernière, Christian; Jouen, Emmanuel; Ah-You, Nathalie; Lefeuvre, Pierre; Chiroleu, Frédéric; Gagnevin, Lionel; Pruvost, Olivier

    2010-03-01

    Three pathogenic variants (i.e. pathotypes) have been described within Xanthomonas citri pv. citri, the causal agent of Asiatic citrus canker. Pathotype A strains naturally infect a wide range of Citrus species and members of some related genera. In contrast, pathotypes A* and A(w) have narrow host ranges within the genus Citrus and have been isolated from Mexican lime (Citrus aurantifolia L.) and from Mexican lime and alemow (Citrus macrophylla L.), respectively. We used amplified fragment length polymorphism (AFLP) and multilocus sequence analysis (MLSA) based on four partial housekeeping gene sequences (atpD, dnaK, efp and gyrB ) for the genotypic classification of Xanthomonas citri pv. citri and the poorly characterized citrus pathogen Xanthomonas campestris pv. bilvae. A Mantel test showed that genetic distances derived from AFLP and MLSA were highly correlated. X. campestris pv. bilvae showed a close relatedness to the type strain of X. citri, indicating that this pathovar should be reclassified as X. citri pv. bilvae. All pathotype A* and A(w) strains were most closely related to X. citri pv. citri strains with a wide host range (pathotype A), confirming previous DNA-DNA hybridization data. Pathotype A(w) should be considered a junior synonym of pathotype A* on the basis of pathogenicity tests, AFLP, MLSA and PCR using pathovar-specific primers. Evolutionary genome divergences computed from AFLP data suggested that pathotype A* (including A(w) strains) is a group of strains that shows a wider genetic diversity than pathotype A.

  10. The small and large subunits of carbamoyl-phosphate synthase exhibit diverse contributions to pathogenicity in Xanthomonas citri subsp. citri

    Institute of Scientific and Technical Information of China (English)

    Guo Jing; SonG Xue; Zou Li-fang; Zou Hua-song; CHen Gong-you

    2015-01-01

    Carbamoyl-phosphate synthase plays a vital role in the carbon and nitrogen metabolism cycles. In Xanthomonas citri subsp. citri, carA and carB encode the smal and large subunits of carbamoyl-phosphate synthase, respectively. The deletion mutation of the coding regions revealed that carA did not affect any of the phenotypes, while carB played multiple roles in pathogenicity. The deletion of carB rendered the loss of pathogenicity in host plants and the ability to induce a hyper-sensitive reaction in the non-hosts. Quantitative reverse transcription-PCR assays indicated that 11 hrp genes coding the type III secretion system were suppressed when interacting with citrus plants. The mutation in carB also affected bacterial utilization of several carbon and nitrogen resources in minimal medium MMX and extracel ular enzyme activities. These data demonstrated that only the large subunit of carbamoyl-phosphate synthase was essential for canker development by X. citri subsp. citri.

  11. Comparative ultrastructure of nonwounded Mexican lime and Yuzu leaves infected with the citrus canker bacterium Xanthomonas citri pv. citri.

    Science.gov (United States)

    Lee, In Jung; Kim, Ki Woo; Hyun, Jae Wook; Lee, Yong Hoon; Park, Eun Woo

    2009-07-01

    Ultrastructural aspects of citrus canker development were investigated in nonwounded leaves of citrus species by transmission electron microscopy (TEM). A susceptible species Mexican lime and a resistant species Yuzu were spray-inoculated with a virulent strain of Xanthomonas citri pv. citri. Initial symptoms occurred on Mexican lime approximately 9 days after inoculation, whereas they appeared on Yuzu mostly 11 days after inoculation. In Mexican lime leaves, the bacterial invasion was usually accompanied by host cell wall dissolution and cellular disruption. Fibrillar materials from degenerated cell walls were usually found in intercellular spaces. Damaged host cells with necrotic cytoplasm showed the localized separation of plasma membrane from the cell wall. Bacterial multiplication and electron-transparent capsule-like structures around bacteria were commonly observed. Meanwhile, cell wall protuberances were prominent outside host cell walls in response to bacterial invasion in Yuzu leaves. Occlusion of intercellular spaces was also formed by the fusion of two or more individual cell wall protuberances originated from two adjacent host cells. Papillae-like materials accumulated locally within host cells in close proximity to bacteria. Some bacteria were found to be undergoing degeneration in xylem vessels. Also, the shrunken, inactive bacteria were surrounded by electron-translucent fibrillar materials in intercellular spaces, implying bacterial immobilization. These cellular responses are thought to be the consequences of defense responses of Yuzu leaves to invading bacteria. In both citrus species, X. citri pv. citri contained polyphosphate bodies showing electron-dense and elliptical structures in cytoplasm.

  12. Unravelling potential virulence factor candidates in Xanthomonas citri. subsp. citri by secretome analysis

    Directory of Open Access Journals (Sweden)

    Rafael M. Ferreira

    2016-02-01

    Full Text Available Citrus canker is a major disease affecting citrus production in Brazil. It’s mainly caused by Xanthomonas citri subsp. citri strain 306 pathotype A (Xac. We analysed the differential expression of proteins secreted by wild type Xac and an asymptomatic mutant for hrpB4 (ΔhrpB4 grown in Nutrient Broth (NB and a medium mimicking growth conditions in the plant (XAM1. This allowed the identification of 55 secreted proteins, of which 37 were secreted by both strains when cultured in XAM1. In this secreted protein repertoire, the following stand out: Virk, Polyphosphate-selective porin, Cellulase, Endoglucanase, Histone-like protein, Ribosomal proteins, five hypothetical proteins expressed only in the wild type strain, Lytic murein transglycosylase, Lipoprotein, Leucyl-tRNA synthetase, Co-chaperonin, Toluene tolerance, C-type cytochrome biogenesis membrane protein, Aminopeptidase and two hypothetical proteins expressed only in the ΔhrpB4 mutant. Furthermore, Peptidoglycan-associated outer membrane protein, Regulator of pathogenicity factor, Outer membrane proteins, Endopolygalacturonase, Chorismate mutase, Peptidyl-prolyl cis-trans isomerase and seven hypothetical proteins were detected in both strains, suggesting that there was no relationship with the secretion mediated by the type III secretory system, which is not functional in the mutant strain. Also worth mentioning is the Elongation factor Tu (EF-Tu, expressed only the wild type strain, and Type IV pilus assembly protein, Flagellin (FliC and Flagellar hook-associated protein, identified in the wild-type strain secretome when grown only in NB. Noteworthy, that FliC, EF-Tu are classically characterized as PAMPs (Pathogen-associated molecular patterns, responsible for a PAMP-triggered immunity response. Therefore, our results highlight proteins potentially involved with the virulence. Overall, we conclude that the use of secretome data is a valuable approach that may bring more knowledge of the

  13. Improved real-time PCR diagnosis of citrus stubborn disease by targeting prophage genes of Spiroplasma citri

    Science.gov (United States)

    Spiroplasma citri is a phloem-limited bacterium causing citrus stubborn disease (CSD). Isolation and culturing of S. citri is difficult and time consuming. Current detection methods use polymerase chain reaction (PCR) assays with primers developed from sequences of S. citri house-keeping genes. In c...

  14. Use of micro-CT to elucidate details of the anatomy and feeding of the Asian citrus psyllid Diaphorina citri Kuwayama, 1908 (Insecta: Hemiptera, Liviidae)

    Science.gov (United States)

    Huanglongbing (HLB), also known as citrus greening disease, is caused by plant-infecting bacteria. The most prominent pathogen within the Americas: United States of America, Mexico, and Brazil, is Candidatus Liberibacter asiaticus, which affects plants of the Family: Rutaceae, in particularly citrus...

  15. Association of ‘Candidatus Liberibacter asiaticus’, the causal agent of citrus huanglongbing in Murraya paniculata and Diaphorina citri in Thailand

    Science.gov (United States)

    Orange jasmine, Murraya paniculata, is a preferred alternative host for the Asian citrus psyllid, the primary vector of citrus Huanglongbing (HLB) disease caused by ‘Candidatus Liberibacter asiaticus’ (Las). M. paniculata plant samples and psyllids on the Murraya plants from ten diverse geographical...

  16. Diversity of the citrus HLB bacterium, ‘Candidatus Liberibacter asiaticus’, in psyllids (Diaphorina citri) collected from Murraya paniculata and citrus spp. in Thailand

    Science.gov (United States)

    ‘Candidatus Liberibacter asiaticus’ (Las) is a phloem inhabiting bacterium that causes huanglongbing disease (HLB), also known as citrus greening associated with three species of a-Proteobacteria in the genus ‘Candidatus Liberibacter sp’. Prophage is an important genetic element of bacterial genomes...

  17. Compatibility of Isaria fumosorosea (Hypocreales: Cordycipitaceae Blastospores with Agricultural Chemicals Used for Management of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae

    Directory of Open Access Journals (Sweden)

    Pasco B. Avery

    2013-11-01

    Full Text Available Biorational insecticides are being increasingly emphasized for inclusion in integrated pest management programs for invasive insects. The entomopathogenic fungus, Isaria fumosorosea, can be used to help manage the Asian citrus psyllid with minimal impact on beneficial arthropods, but its effectiveness may be compromised by agrochemicals used to control concurrent arthropod pests and diseases. We evaluated the compatibility of I. fumosorosea blastospores with a range of spray oils and copper-based fungicides registered for use in citrus groves. Results of laboratory and greenhouse tests showed a range of responses of the fungus to the different materials, including compatibility and incompatibility. Overall, I. fumosorosea growth in vitro was reduced least by petroleum-based materials and most by botanical oils and borax, and some of the copper-based fungicides, suggesting that tank mixing of I. fumosorosea with these latter products should be avoided. However, equivalent negative effects of test materials on fungal pathogenicity were not always observed in tests with adult psyllids. We hypothesize that some oils enhanced adherence of blastospores to the insect cuticle, overcoming negative impacts on germination. Our data show that care should be taken in selecting appropriate agrochemicals for tank-mixing with commercial formulations of entomopathogenic fungi for management of citrus pests. The prospects of using I. fumosorosea for managing the invasive Asian citrus psyllid and other citrus pests are discussed.

  18. First report of citrus canker caused by Xanthomonas citri in Somalia

    Science.gov (United States)

    Xanthomonas citri, causal agent of citrus canker, has been reported in several countries in Africa, but not Somalia. During 2006 and 2007, hyperplasia-type lesions, often surrounded by a water-soaked margin and yellow halo, typical of citrus canker caused by X. citri, were found on 8-10 year-old gr...

  19. Comparison of Droplet Digital PCR and Quantitative PCR Assays for Quantitative Detection of Xanthomonas citri Subsp. citri.

    Directory of Open Access Journals (Sweden)

    Yun Zhao

    Full Text Available Droplet digital polymerase chain reaction (ddPCR is a novel molecular biology technique providing absolute quantification of target nucleic acids without the need for an external calibrator. Despite its emerging applications in medical diagnosis, there are few reports of its use for the detection of plant pathogens. This work was designed to assess the diagnosis potential of the ddPCR for absolute quantitative detection of Xanthomonas citri subsp. citri, a quarantine plant pathogenic bacterium that causes citrus bacterial canker in susceptible Citrus species. We transferred an established quantitative PCR (qPCR assay for citrus bacterial canker diagnosis directly to the ddPCR format and compared the performance of the two methods. The qPCR assay has a broader dynamic range compared to the ddPCR assay and the ddPCR assay has a significantly higher degree of sensitivity compared to the qPCR assay. The influence of PCR inhibitors can be reduced considerably in the ddPCR assay because the collection of end-point fluorescent signals and the counting of binomial events (positive or negative droplets are associated with a Poisson algorithm. The ddPCR assay also shows lower coefficient of variation compared to the qPCR assay especially in low target concentration. The linear association of the measurements by ddPCR and qPCR assays is strong (Pearson correlation = 0.8633; P<0.001. Receiver operating characteristic analysis indicates the ddPCR methodology is a more robust approach for diagnosis of citrus bacterial canker. In summary, the results demonstrated that the ddPCR assay has the potential for the quantitative detection of X. citri subsp. citri with high precision and accuracy as compared with the results from qPCR assay. Further studies are required to evaluate and validate the value of ddPCR technology in the diagnosis of plant disease and quarantine applications.

  20. Morphogenesis of galls induced by Baccharopelma dracunculifoliae (Hemiptera: Psyllidae on Baccharis dracunculifolia (Asteraceae leaves

    Directory of Open Access Journals (Sweden)

    M. Arduin

    Full Text Available The commonest insect gall on Baccharis dracunculifolia (Asteraceae leaves is induced by Baccharopelma dracunculifoliae (Hemiptera, Psyllidae. The gall-inducing insect attacks young leaves in both the unfolded and the fully expanded stages. Four developmental phases were observed in this type of gall: 1 A folding phase, during which the leaf lamina folded upward alongside the midrib and the edges of the upper portion of the leaf approached each other, forming a longitudinal slit. A single chamber was formed on the adaxial surface of the leaf; 2 A swelling phase, in which the folded leaf tissues thickened and the edges of the leaf drew closer together, narrowing the slit. In this phase the gall matured, turning succulent, fusiform and pale green. The single nymphal chamber was lined with white wax and was able to house from one to several nymphs; 3 A dehiscence phase, characterized by the opening of the slit to release inducers; and 4 A senescence phase, when the gall turned dark and dry. The dermal system of the mature gall was composed of a single-layered epidermis. The mesophyll was swollen, and the swelling was due mainly to hyperplasia of the parenchyma. The vascular tissues along the midrib vein were conspicuous and the perivascular fibers resembled parenchymal cells. The hypertrophied secretory cavities contained low lipophylic content. This gall does not form nutritive tissue, but salivary sheaths left by the inducers were observed near the parenchyma, vascular bundles and secretory cavities. This study complements our current knowledge of gall biology and sheds further light on the plasticity of plant tissues stimulated by biotic factors.

  1. Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri.

    Science.gov (United States)

    Zhuo, Tao; Rou, Wei; Song, Xue; Guo, Jing; Fan, Xiaojing; Kamau, Gicharu Gibson; Zou, Huasong

    2015-10-23

    The carA and carB genes code the small and large subunits of carbamoyl-phosphate synthase (CPS) that responsible for arginine and pyrimidine production. The purpose of this work was to study the gene organization and expression pattern of carAB operon, and the biological functions of carA and carB genes in Xanthomonas citri subsp. citri. RT-PCR method was employed to identify the full length of carAB operon transcript in X. citri subsp. citri. The promoter of carAB operon was predicted and analyzed its activity by fusing a GUS reporter gene. The swimming motility was tested on 0.25% agar NY plates with 1% glucose. Biofilm was measured by cell adhesion to polyvinyl chloride 96-well plate. The results indicated that carAB operon was composed of five gene members carA-orf-carB-greA-rpfE. A single promoter was predicted from the nucleotide sequence upstream of carAB operon, and its sensitivity to glutamic acid, uracil and arginine was confirmed by fusing a GUS reporter gene. Deletion mutagenesis of carB gene resulted in reduced abilities in swimming on soft solid media and in forming biofilm on polystyrene microtiter plates. From these results, we concluded that carAB operon was involved in multiple biological processes in X. citri subsp. citri.

  2. Comparative genomics of 43 strains of Xanthomonas citri pv. citri reveals the evolutionary events giving rise to pathotypes with different host ranges.

    Science.gov (United States)

    Gordon, Jonathan L; Lefeuvre, Pierre; Escalon, Aline; Barbe, Valérie; Cruveiller, Stéphane; Gagnevin, Lionel; Pruvost, Olivier

    2015-12-23

    The identification of factors involved in the host range definition and evolution is a pivotal challenge in the goal to predict and prevent the emergence of plant bacterial disease. To trace the evolution and find molecular differences between three pathotypes of Xanthomonas citri pv. citri that may explain their distinctive host ranges, 42 strains of X. citri pv. citri and one outgroup strain, Xanthomonas citri pv. bilvae were sequenced and compared. The strains from each pathotype form monophyletic clades, with a short branch shared by the A(w) and A pathotypes. Pathotype-specific recombination was detected in seven regions of the alignment. Using Ancestral Character Estimation, 426 SNPs were mapped to the four branches at the base of the A, A*, A(w) and A/A(w) clades. Several genes containing pathotype-specific nonsynonymous mutations have functions related to pathogenicity. The A pathotype is enriched for SNP-containing genes involved in defense mechanisms, while A* is significantly depleted for genes that are involved in transcription. The pathotypes differ by four gene islands that largely coincide with regions of recombination and include genes with a role in virulence. Both A* and A(w) are missing genes involved in defense mechanisms. In contrast to a recent study, we find that there are an extremely small number of pathotype-specific gene presences and absences. The three pathotypes of X. citri pv. citri that differ in their host ranges largely show genomic differences related to recombination, horizontal gene transfer and single nucleotide polymorphism. We detail the phylogenetic relationship of the pathotypes and provide a set of candidate genes involved in pathotype-specific evolutionary events that could explain to the differences in host range and pathogenicity between them.

  3. Acaricidal Activity of Boenninghausenia sessilicarpa Against Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    YANG Hui-zhi; HU Jun-hua; LI Qing; LI Hong-jun; LIU Hao-qiang; YAO Ting-shan; RAN Chun; LEI Hui-de

    2009-01-01

    Acaricidal activity of Boenninghausenia sessilicarpa against Panonychus citri was tested in the laboratory.Four solvents were used to prepare crude extracts,petroleum ether,chloroform,ethyl acetate,and ethanol,among which ethanol was the most effective one.The results suggested that ethanol extracts of B.sessilicarpa had eminent acaricidal and ovicidal activities.Concentrated extracts were prepared using petroleum ether,chloroform,ethyl acetate,or distilled water as solvent.Mite mortality rates in the concentrated extracts by petroleum ether,chloroform,or distilled water were significantly lower than those by ethyl acetate.The LC50 values of eggs and female mites were 0.7639 and 1.1033 mg mL-1,respectively.After liquid chromatography and thin layer chromatography,the concentrated extracts were separated into 14 groups of fractions and further tests for their acaricidal and ovicidal activities were conducted.Fraction 2 was found to possess higher acaricidal and ovicidal activities.The mortality of eggs and adult mites were 85.83 and 63.07%,respectively.Moreover,fraction 2 showed moderate oviposition inhibition effect (0.8795) against P.citri when the used dose Was higher than 2.5 mg mL-1.

  4. Different transcriptional response to Xanthomonas citri subsp. citri between kumquat and sweet orange with contrasting canker tolerance.

    Directory of Open Access Journals (Sweden)

    Xing-Zheng Fu

    Full Text Available Citrus canker disease caused by Xanthomonas citri subsp. citri (Xcc is one of the most devastating biotic stresses affecting the citrus industry. Meiwa kumquat (Fortunella crassifolia is canker-resistant, while Newhall navel orange (Citrus sinensis Osbeck is canker-sensitive. To understand the molecular mechanisms underlying the differences in responses to Xcc, transcriptomic profiles of these two genotypes following Xcc attack were compared by using the Affymetrix citrus genome GeneChip. A total of 794 and 1324 differentially expressed genes (DEGs were identified as canker-responsive genes in Meiwa and Newhall, respectively. Of these, 230 genes were expressed in common between both genotypes, while 564 and 1094 genes were only significantly expressed in either Meiwa or Newhall. Gene ontology (GO annotation and Singular Enrichment Analysis (SEA of the DEGs showed that genes related to the cell wall and polysaccharide metabolism were induced for basic defense in both Meiwa and Newhall, such as chitinase, glucanase and thaumatin-like protein. Moreover, apart from inducing basic defense, Meiwa showed specially upregulated expression of several genes involved in the response to biotic stimulus, defense response, and cation binding as comparing with Newhall. And in Newhall, abundant photosynthesis-related genes were significantly down-regulated, which may be in order to ensure the basic defense. This study revealed different molecular responses to canker disease in Meiwa and Newhall, affording insight into the response to canker and providing valuable information for the identification of potential genes for engineering canker tolerance in the future.

  5. Effect of X-irradiation on Citrus Canker Pathogen Xanthomonas citri subsp. citri of Satsuma Mandarin Fruits

    Directory of Open Access Journals (Sweden)

    Min-A Song

    2015-12-01

    Full Text Available Citrus canker caused by Xanthomonas citri subsp. citri (Xcc is one of the most important bacterial diseases of citrus. Because citrus canker is not found in many countries including European Union and Australia, Xcc is strictly regulated in order to prevent its spread. In this study, the effects of X-irradiation on Xcc growth either in the suspension or on the surface of citrus fruits were investigated. The suspension containing 1×10⁷ cfu/ml of Xcc was irradiated with different absorbed doses of X-irradiation ranging from 50 to 400 Gy. The results showed that Xcc was fully dead at 400 Gy of X-irradiation. To determine the effect of X-irradiation on quarantine, the Xcc-inoculated citrus fruits were irradiated with different X-ray doses at which Xcc was completely inhibited by an irradiation dose of 250 Gy. The D₁₀ value for Xcc on citrus fruits was found to be 97 Gy, indicating the possibility of direct application on citrus quarantine without any side sterilizer. Beside, presence of Xcc on the surface of asymptomatic citrus fruits obtained from citrus canker-infected orchards was noted. It indicated that the exporting citrus fruits need any treatment so that Xcc on the citrus fruits should be completely eliminated. Based on these results, ionizing radiation can be considered as an alternative method of eradicating Xcc for export of citrus fruits.

  6. Closely-related Xanthomonas citri subsp. citri isolates trigger distinct histological and transcriptional responses in Citrus limon

    Directory of Open Access Journals (Sweden)

    Ingrid Georgina Orce

    Full Text Available ABSTRACT Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc, has an important economic impact on the citrus industry. Extensive information is available about the disease but, nevertheless, the study of plant-pathogen interactions could provide new information in the understanding of citrus canker disease. A new isolate has been identified, Xcc AT, which has a high genetic similarity (> 90 % to the virulent Xcc T strain based on genetic clustering analyses of the rep-PCR fingerprinting patterns, but it does not produce cankerous lesions in Citrus limon. In this study, we compared C. limon responses to Xcc AT and to the virulent Xcc T strain at both histological and transcriptional levels. Histologically, leaves inoculated with Xcc AT exhibited neither a typical disordering of the spongy mesophyll, nor a swelling of epidermis. A particular content (undetermined was also found in mesophyll cells near the stomata, together with increased starch accumulation. The transcriptomic profiles were compared by cDNA-AFLP technique. A total of 121 fragments derived from transcript (TDF were either specifically induced or repressed by the isolates, and 62 were sequenced. Analysis of global expression identified different classes of genes known to be involved in plant-pathogen interactions. This study constitutes the first approach of the specific interaction between the avirulent Xcc AT isolate and C. limon.

  7. The role as inoculum sources of Xanthomonas citri pv. citri surviving on the infected Satsuma mandarin fruits.

    Science.gov (United States)

    Kang, So Young; Kim, Ki Deok; Hong, Jeum Kyu; Hyun, He Nam; Jeun, Yong Chull

    2014-05-01

    Importing citrus fruits infected by Asiatic citrus canker caused by Xanthomonas citri pv. citri (Xcc) can act as an inoculum source for the disease epidemic in citrus canker-free countries. In this study, the pathogenicity of the causal agent of Asiatic citrus canker surviving on infected Satsuma mandarin fruits was evaluated. The washing solution of infected Satsuma mandarin fruits did not cause lesion formation on the citrus leaves. However, a typical citrus canker lesion was formed on the leaves after inoculation with higher concentrations of the inoculum from the washing solution (washing solution II). It indicated that the pathogenicity of the citrus canker surviving on the symptomatic Satsuma mandarin fruits was not changed. Scanning electron microscopic observation showed that the numbers of bacterial cells on the leaves of Satsuma mandarin which inoculated with the washing solution directly (washing solution I) was less compared to those of leaves inoculated with the washing solution II. This result spports that the pathogenicity of Xcc surviving on Satsuma mandarin fruits may not be changed but that the sucessful infection of citrus caker may depend on the concentration of the inoculum.

  8. Use of recombinant tobacco mosaic virus to achieve RNA interference in plants against the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae.

    Directory of Open Access Journals (Sweden)

    Arif Muhammad Khan

    Full Text Available The citrus mealybug, Planococcus citri, is an important plant pest with a very broad plant host range. P. citri is a phloem feeder and loss of plant vigor and stunting are characteristic symptoms induced on a range of host plants, but P. citri also reduces fruit quality and causes fruit drop leading to significant yield reductions. Better strategies for managing this pest are greatly needed. RNA interference (RNAi is an emerging tool for functional genomics studies and is being investigated as a practical tool for highly targeted insect control. Here we investigated whether RNAi effects can be induced in P. citri and whether candidate mRNAs could be identified as possible targets for RNAi-based P. citri control. RNAi effects were induced in P. citri, as demonstrated by specific target reductions of P. citri actin, chitin synthase 1 and V-ATPase mRNAs after injection of the corresponding specific double-stranded RNA inducers. We also used recombinant Tobacco mosaic virus (TMV to express these RNAi effectors in Nicotiana benthamiana plants. We found that P. citri showed lower fecundity and pronounced death of crawlers after feeding on recombinant TMV-infected plants. Taken together, our data show that actin, chitin synthase 1 and V-ATPase mRNAs are potential targets for RNAi against P. citri, and that recombinant TMV is an effective tool for evaluating candidate RNAi effectors in plants.

  9. Use of recombinant tobacco mosaic virus to achieve RNA interference in plants against the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae).

    Science.gov (United States)

    Khan, Arif Muhammad; Ashfaq, Muhammad; Kiss, Zsofia; Khan, Azhar Abbas; Mansoor, Shahid; Falk, Bryce W

    2013-01-01

    The citrus mealybug, Planococcus citri, is an important plant pest with a very broad plant host range. P. citri is a phloem feeder and loss of plant vigor and stunting are characteristic symptoms induced on a range of host plants, but P. citri also reduces fruit quality and causes fruit drop leading to significant yield reductions. Better strategies for managing this pest are greatly needed. RNA interference (RNAi) is an emerging tool for functional genomics studies and is being investigated as a practical tool for highly targeted insect control. Here we investigated whether RNAi effects can be induced in P. citri and whether candidate mRNAs could be identified as possible targets for RNAi-based P. citri control. RNAi effects were induced in P. citri, as demonstrated by specific target reductions of P. citri actin, chitin synthase 1 and V-ATPase mRNAs after injection of the corresponding specific double-stranded RNA inducers. We also used recombinant Tobacco mosaic virus (TMV) to express these RNAi effectors in Nicotiana benthamiana plants. We found that P. citri showed lower fecundity and pronounced death of crawlers after feeding on recombinant TMV-infected plants. Taken together, our data show that actin, chitin synthase 1 and V-ATPase mRNAs are potential targets for RNAi against P. citri, and that recombinant TMV is an effective tool for evaluating candidate RNAi effectors in plants.

  10. The push-pull strategy for citrus psyllid control.

    Science.gov (United States)

    Yan, Huaxue; Zeng, Jiwu; Zhong, Guangyan

    2015-07-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the only natural vector of Candidatus Liberibacter asiaticus that causes citrus huanglongbing (HLB), a most destructive disease of citrus. Currently, no remedial therapy exists for the disease, and so effective control of ACP is very important in curbing the transmission of the disease. The push-pull strategy should be thoroughly explored as an approach to ACP management. This mini-review summarises the current progress towards more effective repellent and attractant chemicals through investigating known repellent and attractive plants. Interactions between ACP and its host plants are also addressed, with emphasis on the possible involvement of the host biochemicals in attracting the insect. Potential ways to increase the effectiveness of the pull-push strategy are briefly discussed. It is expected that the pull-push strategy will be gradually developed following more extensive research.

  11. [Analysis on medication rules of state medical master Yan Zhenghua from prescriptions with citri reticulatae pericarpium based on data mining].

    Science.gov (United States)

    Wu, Jia-Rui; Guo, Wei-Xian; Zhang, Bing; Zhang, Xiao-Meng; Yang, Bing; Sheng, Xiao-Guang

    2014-02-01

    The prescriptions containing pericarpium citri reticulatae that built by Professor. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. After analyzed by data mining, such as apriori algorithm, the frequency of single medicine, the frequency of drug combination, the association rules between drugs and core drug combinations can be get from the database. Through the analysis of 1 027 prescriptions with pericarpium citri reticulatae, these prescriptions were commonly used to treat stomach aches, cough and other syndromes. The most frequency drug combinations were "Citri Reticulatae Pericarpium-Poria", "Paeoniae Radix Rubra-Citri Reticulatae Pericarpium" and so on. The drug association rules that the confidence was 1 were "Glycyrrhizae Radix ex Rhizoma --> Citri Reticulatae Pericarpium", "Paeoniae Alba Radix-Cyperi Rhizoma --> Citri Reticulatae Pericarpium", "Poria --> Citri Reticulatae Pericarpium", and so on. The drugs in the prescriptions containing pericarpium citri reticulatae that built by Professor Yan mostly had the effects of regulating the flow of Qi and invigorate blood circulation, which reflected the clearly thought when making prescriptions.

  12. Energy allocation changes in overwintering adults of the common pistachio Psylla, Agonoscena pistaciae Burckhardt & Lauterer (Hemiptera: Psyllidae).

    Science.gov (United States)

    Sadeghi, R; Izadi, H; Mahdian, K

    2012-12-01

    The common pistachio psylla, Agonoscena pistaciae Burckhardt & Lauterer (Hemiptera: Psyllidae), is known as the key pest of pistachio orchards in Iran. This pest passes the winter as adults. In this study, energy allocation changes in relation to ambient temperature were investigated in field-collected adults by measuring total body sugar, trehalose, glucose, sorbitol, myoinositol, glycogen, lipid, and protein contents. Glycogen content decreased with decrease in ambient temperature. The decrease in glycogen content was proportional to the increase in total body sugar, trehalose, myoinositol, and sorbitol contents. In January, with mean ambient temperature of 5.4°C, glycogen content was at the lowest level, whereas total body sugar, trehalose, glucose, and sorbitol were at the highest level. Total body sugar, trehalose, myoinositol, and sorbitol contents increased as temperature decreased from 22.7°C in October to 5.4°C in January. In conclusion, low molecular weight carbohydrates and polyols may play a role in winter survival and adaptation to cold of the common pistachio psylla by providing the required cryoprotection. Also, overwintering adults of the common pistachio psylla may store energy in the form of lipid for later utilization during the overwintering.

  13. Testing Spirotetramat as an Alternative Solution to Abamectin for Cacopsylla pyri (Hemiptera: Psyllidae) Control: Laboratory and Field Tests.

    Science.gov (United States)

    Civolani, Stefano; Boselli, Mauro; Butturini, Alda; Chicca, Milvia; Cassanelli, Stefano; Tommasini, Maria Grazia; Aschonitis, Vassilis; Fano, Elisa Anna

    2015-12-01

    Aim of the study was to investigate the performance of the new insecticide "spirotetramat" as an alternative solution of "abamectin" for the control of Cacopsylla pyri L. (Hemiptera: Psyllidae) in the context of an IPM program in European pear, Pyrus communis L.. Laboratory bioassays for the estimation of LC50 and LC90 of both insecticides were performed using four populations collected in Emilia-Romagna (Italy) orchards where different pest management strategies were used (organic, integrated, and conventional). The same populations were also analyzed for the main insecticide detoxifying activities in nymphs by spectrofluorimetric in vitro assays. The performance of the two insecticides was also tested on field on one population under integrated pest management conditions. The laboratory experiments showed that the LC90 of spirotetramat were lower than the highest field concentration allowed in Europe (172.80 mg AI liter(-1)) giving reassurance about the efficacy of the product. Concerning the abamectin, the laboratory bioassays did not show strong indications of resistance development of C. pyri populations of Emilia-Romagna. A similarity in enzyme detoxifying activity was observed in both insecticides indicating a general absence of a significant insecticide resistance. The field trial showed a high efficacy (>90 %) of spirotetramat on C. pyri already after 15 d from application, and it was significantly higher from abamectin. Overall, spirotetramat is one more choice for C. pyri control, as well as abamectin in order to minimize the risks of occurrence of insecticide resistance.

  14. A Virulence-Reducing Mutation in the Postharvest Citrus Pathogen Alternaria citri.

    Science.gov (United States)

    Katoh, H; Isshiki, A; Masunaka, A; Yamamoto, H; Akimitsu, K

    2006-09-01

    ABSTRACT Alternaria citri causes Alternaria black rot, a postharvest fruit disease, on a broad range of citrus cultivars. We previously described that an endopolygalacturonase minus mutant of A. citri caused significantly less black rot in citrus fruit. To search for other essential factors causing symptoms in addition to endopolygalacturonase, a random mutation analysis of pathogenicity was performed using restriction enzyme-mediated integration. Three isolates among 1,694 transformants of A. citri had a loss in pathogenicity in a citrus peel assay, and one of these three mutants was a histidine auxotroph. Gene AcIGPD that encodes imidazole glycerol phosphate dehydratase, the sixth enzyme in the histidine biosynthetic pathway, was cloned, and the mutant containing the disrupted target gene, AcIGPD, caused less black rot.

  15. Characterization and activity of entomopathogenic fungi isolates against “paraguay tea ampul” (Gyropsylla spegazziniana (Lizer & Trelles (Hemiptera: Psyllidae

    Directory of Open Access Journals (Sweden)

    Marina Andressa Formentini

    2015-12-01

    Full Text Available Paraguay tea is a native crop of southern Brazil and is also socioeconomically important in Paraguay and Argentina. One of the main pests of this crop is Gyropsylla spegazziniana (Hemiptera: Psyllidae. As there are no registered insecticides for this pest in Brazil, the use of entomopathogenic fungi is an alternative method for its control. This study aimed to evaluate and characterize isolates of the entomopathogenic fungi Beauveria spp., Metarhizium anisopliae, Isaria spp. and Lecanicillium spp. on G. spegazziniana. For this purpose, 5th instar nymphs were transferred to Paraguay tea seedlings, followed by spraying of conidial suspensions (1 × 109 conidia mL-1 and then placing the seedlings in polyvinyl chloride (PVC cages and maintaining them in a temperature-controlled room (26 ± 1°C, 12h00 photophase, 60 ± 10% R.H.. Insect mortality was evaluated daily for 10 days, and the selected isolates were characterized according to their vegetative growth, conidial production in synthetic culture medium and rice and insecticidal activity as well as through molecular analyses, including sequencing the rDNA-ITS region and RAPD analysis. The genus Beauveria spp. was the most efficient, particularly the Unioeste 44 isolate, which caused the greatest total mortality of G. spegazziniana (81.7% and showed among the highest conidial production levels on rice, indicating a significant potential to be used in an integrated management program for this pest. Molecular analysis of the rDNA-ITS region allowed the isolates to be identified as B. bassiana and B. brongniartii, and RAPD markers were found to be associated with virulence.

  16. Molecular Detection of Spiroplasma Citri Associated with Stubborn Disease in Citrus Orchards in Syria

    Science.gov (United States)

    Spiroplasma citri, a phloem-limited pathogen, causes citrus stubborn disease (CSD) and can be transmitted from plant to plant by several species of phloem-feeding leafhoppers. CSD is an important disorder in certain warm and arid citrus-growing areas, and its agent has been recorded from several Med...

  17. Geïntegreerde bestrijding van citruswolluis Planococcus citri in roos

    NARCIS (Netherlands)

    Pijnakker, J.; Leman, A.; Staaij, van der M.

    2011-01-01

    Referaat Sinds begin 2000 is citruswolluis, Planococcus citri (Risso), een van de belangrijkste plagen geworden in de rozenteelt onder glas. Telers kiezen voor een meer geïntegreerde aanpak om andere plagen onder controle te houden en stoppen met het reguliere gebruik van breedwerkende middelen. Hie

  18. Geïntegreerde bestrijding van citruswolluis Planococcus citri in roos

    NARCIS (Netherlands)

    Pijnakker, J.; Leman, A.; Hennekam, M.

    2013-01-01

    Citruswolluis, Planococcus citri (Risso), is een van de belangrijkste plagen geworden in de rozenteelt onder glas. Er is inmiddels veel praktijkervaring met de toepassing van biologische bestrijders om haarden uit te roeien. Voor de professionele tuinbouw is de effectiviteit van deze aanpak doorgaan

  19. Reduced Susceptibility to Xanthomonas citri in Transgenic Citrus Expressing the FLS2 Receptor From Nicotiana benthamiana.

    Science.gov (United States)

    Hao, Guixia; Pitino, Marco; Duan, Yongping; Stover, Ed

    2016-02-01

    Overexpression of plant pattern-recognition receptors by genetic engineering provides a novel approach to enhance plant immunity and broad-spectrum disease resistance. Citrus canker disease associated with Xanthomonas citri is one of the most important diseases damaging citrus production worldwide. In this study, we cloned the FLS2 gene from Nicotiana benthamiana cDNA and inserted it into the binary vector pBinPlus/ARS to transform Hamlin sweet orange and Carrizo citrange. Transgene presence was confirmed by polymerase chain reaction (PCR) and gene expression of NbFLS2 was compared by reverse transcription quantitative PCR. Reactive oxygen species (ROS) production in response to flg22Xcc was detected in transgenic Hamlin but not in nontransformed controls. Low or no ROS production was detected from nontransformed Hamlin seedlings challenged with flg22Xcc. Transgenic plants highly expressing NbFLS2 were selected and were evaluated for resistance to canker incited by X. citri 3213. Our results showed that the integration and expression of the NbFLS2 gene in citrus can increase canker resistance and defense-associated gene expression when challenged with X. citri. These results suggest that canker-susceptible Citrus genotypes lack strong basal defense induced by X. citri flagellin and the resistance of these genotypes can be enhanced by transgenic expression of the flagellin receptor from a resistant species.

  20. Antifungal Effect of the Aqueous and Ethanolic Avicennia marina Extracts on Alternaria citri and Penicillium digitatum

    Directory of Open Access Journals (Sweden)

    Behrooz Alizadeh Behbahani

    2016-02-01

    Full Text Available Background Plants have a great potential for producing new drugs of great benefit to mankind. There are many approaches to the search for new biologically active principles in higher plants. Objectives The aim of this study was to determine antifungal effect of the aqueous and ethanolic extracts of Avicennia marina on Penicillium digitatum and Alternaria citri in vitro. Materials and Methods In this experimental study, antimicrobial effects of extract were determined by screening antimicrobial activity and disk agar diffusion test in 20, 40, 60 and 80 mg/mL concentration of the extract against P. digitatum and A. citri. Minimum inhibitory concentration (MIC and minimum fungicidal concentration (MFC were determined by incorporating various concentrations of extracts in Sabouraud dextrose agar (SDA in tubes against P. digitatum and A. citri. The data were analyzed using one way analysis of variance (ANOVA using SPSS-17. Results The results indicate that extracts of A. marina mostly had been effective on P. digitatum and have the least impact on A. citri. The results show that the ethanol extract of mangrove leaf with antimicrobial activity method in 2 mg/mL, were inhibited the growth of P. digitatum and A. citri. In disk agar diffusion test the mentioned extract were shown inhibition effect on pathogen molds growth (P < 0.05. Conclusions Ethanolic extract compared to the aqueous extract was more effective and has a greater inhibition effect. The results of this study showed that the extract of A. marina has antifungal properties that justify their traditional use as medicinal plants.

  1. Expressão gênica diferencial de laranja Pêra Rio (Citrus sinensis (L.) Osbeck) e Lima Ácida 'Galego' (Citrus aurantifolia Swingle) em resposta à infecção por Xanthomonas citri subsp. citri

    OpenAIRE

    Cavallini, Juliana da Silva [UNESP

    2013-01-01

    The citrus agribusiness is very important to the Brazilian economy, but the increase of diseases in the last decade has caused great economic losses to the sector. The citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (Xac), is a serious disease that attacks all citrus species economically important worldwide and there is not an effective method for its control. In this study, RNASeq was used to analyze the transcriptional profiles of two contrasting citrus genotypes regar...

  2. Sexual transmission of a plant pathogenic bacterium, Candidatus Liberibacter asiaticus, between conspecific insect vectors during mating.

    Directory of Open Access Journals (Sweden)

    Rajinder S Mann

    Full Text Available Candidatus Liberibacter asiaticus is a fastidious, phloem-inhabiting, gram-negative bacterium transmitted by Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae. The bacterium is the presumed causal agent of huanglongbing (HLB, one of the most destructive and economically important diseases of citrus. We investigated whether Las is transmitted between infected and uninfected D. citri adults during courtship. Our results indicate that Las was sexually transmitted from Las-infected male D. citri to uninfected females at a low rate (<4% during mating. Sexual transmission was not observed following mating of infected females and uninfected males or among adult pairs of the same sex. Las was detected in genitalia of both sexes and also in eggs of infected females. A latent period of 7 days or more was required to detect the bacterium in recipient females. Rod shaped as well as spherical structures resembling Las were observed in ovaries of Las-infected females with transmission electron microscopy, but were absent in ovaries from uninfected D. citri females. The size of the rod shaped structures varied from 0.39 to 0.67 µm in length and 0.19 to 0.39 µm in width. The spherical structures measured from 0.61 to 0.80 µm in diameter. This investigation provides convincing evidence that a plant pathogenic bacterium is sexually transmitted from male to female insects during courtship and established evidence that bacteria persist in reproductive organs. Moreover, these findings provide an alternative sexually horizontal mechanism for the spread of Las within populations of D. citri, even in the absence of infected host trees.

  3. Green Fluorescent Detection of Fungal Colonization and Endopolygalacturonase Gene Expression in the Interaction of Alternaria citri with Citrus.

    Science.gov (United States)

    Isshiki, A; Ohtani, K; Kyo, M; Yamamoto, H; Akimitsu, K

    2003-07-01

    ABSTRACT Alternaria citri, a postharvest pathogen, produces endopolygalacturonase (endoPG) and causes black rot on citrus fruit. We previously described that an endoPG-disrupted mutant of Alternaria citri was significantly reduced in its ability to macerate plant tissue and cause black rot symptoms on citrus. In order to investigate colonization of citrus fruit tissues by Alternaria citri, pTEFEGFP carrying a green fluorescent protein (GFP) gene was introduced into wild-type Alternaria citri and its endoPG-disrupted mutant (M60). Green fluorescence was observed in spores, germ tubes, appressoria, and infection hyphae of transformants G1 (derived from wild type) and GM4 (derived from M60). Hyphae of G1 but not GM4 vertically penetrated the peel, but the hyphae of both G1 and GM4 spread equally in the juice sac area of citrus fruit. Green fluorescence of Alternaria citri transformant EPG7 carrying a GFP gene under control of the endoPG gene promoter of Alternaria citri was induced by pectin in the peel during the infection stage, but repressed completely in the juice sac area, likely by carbon catabolite repression by sugars in the juice.

  4. Biologia de Neotrioza tavaresi Crawford, 1925 (Hemiptera, Psyllidae, galhador da folha do araçazeiro (Psidium cattleianum

    Directory of Open Access Journals (Sweden)

    C. A. Butignol

    2003-01-01

    Full Text Available Biology of the leaf gall inducer Neotrioza tavaresi Crawford, 1925 (Hemiptera, Psyllidae on strawberry guava tree (Psidium cattleianum. A field study was conducted in Curitiba region, State of Paraná, southern Brazil, to describe the life cycle of Neotrioza tavaresi Crawford, 1925, a leaf galling insect in strawberry guava trees (Psidium cattleianum. Three cycles were observed (1997, 1998, 1999 during regular field trips and the insects were observed in Piraquara municipality, where 15 samples with 50 infested leaves were sampled in the 1997-98 cycle. Galls were dissected for detailed studies. Neotrioza tavaresi has a univoltine cycle in which adult individuals were found inside the galls from August onwards. The sexually mature insects with sex ratio 1, emerged from the galls after their dehiscence caused by feeding of the adult insects on the gall walls. Adult emergence started in early October and ended by early December, with its peak in November. Copulation took place as soon as adults exit the gall and egg laying started the next day. Females had more than 100 ovarioles containing 218.7±44.7 (n=50 fully formed eggs. This indicated the short sexual adult life-span (aprox. 5-7 days of the species, also characterized by a concentrated oviposition. Adult individuals fed and laid their eggs on younger shoots of the plant. The bottoms of the yellowish eggs were inserted into the leaf tissue, mainly on its adaxial edge (78.1%. The nymphs hatched and, as they fed on the adaxial side of expanding leaves, modified the cell growth pattern and the round-shape galls developed on the adaxial side with one insect inside. The gall wall showed distinct layers, with the inner one suppliyng the food to the insects, and the outer layer supplying gall protection. Nymphs went through five instars and the exuviae remained stuck on a ball of wax inside the gall. All parasitoids found were Hymenoptera belonging to Chalcidoidea: Eulophidae (1 sp, Pteromalidae (2

  5. Structural and functional analyses of the putrescine binding protein PotF from Xanthomonas citri

    Energy Technology Data Exchange (ETDEWEB)

    Santana, L.D.F.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

    2012-07-01

    Full text: The focus of our group is to determinate the role of ABC transporters in the physiology and growth of Xanthomonas citri, a phytopathogenic bacteria that infects citrus plants causing significant losses for the economy. One of the ABC transporters identified in the X. citri genome and that was showed to be active during the infection in Citrus sinensis plants was the putrescine transporter. This transporter consists of two internal membrane proteins PotG and PotH that form a pore, a cytoplasmic protein that gives energy for the transport and the periplasmic-binding protein PotF, which is responsible for the affinity and specificity of the system. Its function is associated to the microbial carcinogenesis, biofilm formation, escape from phagolysosomes, bacteriocin production, toxin activity and protection from oxidative and acid stress. In this work, we show for the first time, the expression, purification, functional and structural analyses of the X. citri PotF protein. The PotF was expressed from Escherichia coli cells strain Arctic, as a 40 kDa soluble protein, after induction of IPTG for twenty four hours at thirteen deg C. Using immobilized metal affinity chromatography for purification, the protein was eluted in the fractions with 10-500 mM of imidazole. To test the folding and cability to bind putrescine, spectroscopic analyses were performed using circular dichroism and intrinsic fluorescence. The data showed that PotF suffers conformational changes in presence of ligands and in different pH, suggesting a possible interaction with the tested ligand. Moreover, based on bioinformatics studies and molecular modeling analyses, we showed that X. citri PotF is highly conserved when compared to orthologs present in other bacteria, including the residues that form the ligand-binding site. The production of PotF in a soluble and stable form will allow us to start the crystallization trials in attempt to solve its structure. (author)

  6. Novel diagnosis for citrus stubborn disease by detection of a spiroplasma citri-secreted protein.

    Science.gov (United States)

    Shi, Jinxia; Pagliaccia, Deborah; Morgan, Robyn; Qiao, Yongli; Pan, Songqin; Vidalakis, Georgios; Ma, Wenbo

    2014-02-01

    Citrus stubborn disease (CSD), first identified in California, is a widespread bacterial disease found in most arid citrus-producing regions in the United States and the Mediterranean Region. The disease is caused by Spiroplasma citri, an insect-transmitted and phloem-colonizing bacterium. CSD causes significant tree damage resulting in loss of fruit production and quality. Detection of CSD is challenging due to low and fluctuating titer and sporadic distribution of the pathogen in infected trees. In this study, we report the development of a novel diagnostic method for CSD using an S. citri-secreted protein as the detection marker. Microbial pathogens secrete a variety of proteins during infection that can potentially disperse systemically in infected plants with the vascular flow. Therefore, their distribution may not be restricted to the pathogen infection sites and could be used as a biological marker for infection. Using mass spectrometry analysis, we identified a unique secreted protein from S. citri that is highly expressed in the presence of citrus phloem extract. ScCCPP1, an antibody generated against this protein, was able to distinguish S. citri-infected citrus and periwinkle from healthy plants. In addition, the antiserum could be used to detect CSD using a simple direct tissue print assay without the need for sample processing or specialized lab equipment and may be suitable for field surveys. This study provides proof of a novel concept of using pathogen-secreted protein as a marker for diagnosis of a citrus bacterial disease and can probably be applied to other plant diseases.

  7. Adjuvants to improve aerial control of the citrus mealybug Planococcus citri (Hemiptera: Pseudococcidae) using entomopathogenic nematodes.

    Science.gov (United States)

    van Niekerk, S; Malan, A P

    2015-03-01

    The citrus mealybug, Planococcus citri, is a highly destructive pest of citrus, occurring only in the aerial parts of plants. Humidity will be one of the key factors to consider when using entomopathogenic nematodes (EPN) as biological control agents. Different adjuvants can be added to suspensions of EPNs, to improve control as a foliar application. An aqueous suspension containing Heterorhabditis zealandica and 0.3% Zeba® significantly increased P. citri mortality by 22% at 80% relative humidity (RH) with a temperature cycle starting at 22°C for 14 h and 11°C for 11 h. The same polymer formulation was tested for Steinernema yirgalemense and mortality of P. citri increased by 21% at 60% RH and by 27% at 80% RH. The addition of Nu-Film-P® and Zeba® to H. zealandica suspensions did not significantly retard application run-off on citrus leaves. The combination of Nu-Film-P® and Zeba®, however, was able to significantly retard sedimentation, increasing the average number of nematodes deposited on 2-cm2 leaf discs by 10 nematodes. In an aqueous suspension, nematodes settle rapidly to the bottom, resulting in an uneven distribution of nematodes. Xanthan gum, at a concentration of 0.2%, was highly effective at retarding sedimentation, with 72% of the initial nematode number still in suspension after 1 h. Zeba®, at a concentration of 0.3%, despite not being as effective as Xanthan gum, nevertheless still retarded sedimentation significantly. This is the first report of the potential of Nu-Film-P® and Zeba® to improve EPN performance against P. citri when used above ground in citrus orchards.

  8. Formic and acetic acids in degradation products of plant volatiles elicit olfactory and behavorial responses from an insect vector

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae) vectors the bacterial pathogen presumed to be the etiological agent of citrus greening disease, Huanglongbing (HLB), a major threat to citrus industry worldwide. We studied antennal and behavioral responses of Diaphorina citri...

  9. The relevance of Murraya paniculata and related species as potential hosts and inoculum reservoirs of Candidatus Liberibacter asiaticus, causal agent of Huanglongbing (HLB)

    Science.gov (United States)

    Huanglongbing (HLB), considered the most serious insect-vectored bacterial disease of citrus, is transmitted in nature by the Asian citrus psyllid Diaphorina citri and the African citrus psyllid Trioza erytreae. Diaphorina citri was discovered in southern Florida in 1998 and the HLB disease in 2005...

  10. Genetic diversity of Xanthomonas axonopodis pv. citri based on plasmid profile and pulsed field gel electrophoresis

    Directory of Open Access Journals (Sweden)

    Carvalho Flávia Maria de Souza

    2005-01-01

    Full Text Available Xanthomonas axonopodis pv. citri strains that cause disease in citrus were investigated by pulsed field and plasmid profile analysis. For the first method, genomic DNA was digested by the rare-cutting enzymes Xba I and Vsp I. The strains evaluated were collected in seven different States of Brazil and in Argentina, Bolivia, Paraguay and Uruguay. Genetic variability was found among strains of X. axonopodis pv. citri from different geographical areas Argentina, Bolivia and Uruguay, with similarities varying from 0.62 to 0.83. However, the strains collected in Brazil, despite being from different States, have shown a genetic similarity ranging from 0.83 to 1.00. Cluster analysis showed a relationship between genomic similarity and geographical origin of the strains. Plasmids were observed in all strains, with a total of five different plasmids, with sizes between 57.7 and 83.0 kilobases. The 72.6 kb plasmid was the most frequent, present in 15 out of 22 strains, while the 68.1 kb plasmid was observed in two strains only. Although the plasmid diversity detected in the present study was not very great, the X. axonopodis pv. citri strains evaluated showed a considerable degree of diversity with regard to this extrachromosomal genetic element.

  11. Molecular Effects of Irradiation (Cobalt-60 on the Control of Panonychus citri (Acari: Tetranychidae

    Directory of Open Access Journals (Sweden)

    Ke Zhang

    2015-11-01

    Full Text Available The effective dose of irradiation to control pest mites in quarantine has been studied extensively, but the molecular mechanisms underlying the effects of the irradiation on mites are largely unknown. In this study, exposure to 400 Gy of γ rays had significant (p < 0.05 effects on the adult survival, fecundity and egg viability of Panonychus citri. The irradiation caused the degradation of the DNA of P. citri adults and damaged the plasma membrane system of the egg, which led to condensed nucleoli and gathered yolk. Additionally, the transcriptomes and gene expression profiles between irradiated and non-irradiated mites were compared, and three digital gene expression libraries were assembled and analyzed. The differentially expressed genes were putatively involved in apoptosis, cell death and the cell cycle. Finally, the expression profiles of some related genes were studied using quantitative real-time PCR. Our study provides valuable information on the changes in the transcriptome of irradiated P. citri, which will facilitate a better understanding of the molecular mechanisms that cause the sterility induced by irradiation.

  12. The molybdate-binding protein (ModA) of the plant pathogen Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Balan, Andrea; Santacruz, Carolina P; Moutran, Alexandre; Ferreira, Rita C C; Medrano, Francisco J; Pérez, Carlos A; Ramos, Carlos H I; Ferreira, Luís C S

    2006-12-01

    The modABC operon of phytopathogen Xanthomonas axonopodis pv. citri (X. citri) encodes a putative ABC transporter involved in the uptake of the molybdate and tungstate anions. Sequence analyses showed high similarity values of ModA orthologs found in X. campestris pv. campestris (X. campestris) and Escherichia coli. The X. citri modA gene was cloned in pET28a and the recombinant protein, expressed in the E. coli BL21 (DE3) strain, purified by immobilized metal affinity chromatography. The purified protein remained soluble and specifically bound molybdate and tungstate with K(d) 0.29+/-0.12 microM and 0.58+/-0.14 microM, respectively. Additionally binding of molybdate drastically enhanced the thermal stability of the recombinant ModA as compared to the apoprotein. This is the first characterization of a ModA ortholog expressed by a phytopathogen and represents an important tool for functional, biochemical and structural analyses of molybdate transport in Xanthomonas species.

  13. Xanthomonas citri subsp. citri surface proteome by 2D-DIGE: Ferric enterobactin receptor and other outer membrane proteins potentially involved in citric host interaction.

    Science.gov (United States)

    Carnielli, Carolina Moretto; Artier, Juliana; de Oliveira, Julio Cezar Franco; Novo-Mansur, Maria Teresa Marques

    2017-01-16

    Xanthomonas citri subsp. citri (XAC) is the causative agent of citrus canker, a disease of great economic impact around the world. Understanding the role of proteins on XAC cellular surface can provide new insights on pathogen-plant interaction. Surface proteome was performed in XAC grown in vivo (infectious) and in vitro (non-infectious) conditions, by labeling intact cells followed by cellular lysis and direct 2D-DIGE analysis. Seventy-nine differential spots were analyzed by mass spectrometry. Highest relative abundance for in vivo condition was observed for spots containing DnaK protein, 60kDa chaperonin, conserved hypothetical proteins, malate dehydrogenase, phosphomannose isomerase, and ferric enterobactin receptors. Elongation factor Tu, OmpA-related proteins, Oar proteins and some Ton-B dependent receptors were found in spots decreased in vivo. Some proteins identified on XAC's surface in infectious condition and predicted to be cytoplasmic, such as DnaK and 60KDa chaperonin, have also been previously found at cellular surface in other microorganisms. This is the first study on XAC surface proteome and results point to mediation of molecular chaperones in XAC-citrus interaction. The approach utilized here can be applied to other pathogen-host interaction systems and help to achieve new insights in bacterial pathogenicity toward promising targets of biotechnological interest.

  14. Potential of South African entomopathogenic nematodes (Heterorhabditidae and Steinernematidae) for control of the citrus mealybug, Planococcus citri (Pseudococcidae).

    Science.gov (United States)

    van Niekerk, Sonnica; Malan, Antoinette P

    2012-10-01

    Planococcus citri, the citrus mealybug, is the most important species of mealybug known to infest citrus in South Africa. Various laboratory bioassays were conducted to determine the potential of entomopathogenic nematodes to control P. citri. Adult female P. citri were screened for susceptibility to six indigenous nematode species. P. citri was found to be most susceptible to Steinernema yirgalemense and Heterorhabditis zealandica, causing 97% and 91% mortality, respectively. The development of nematodes after infecting adult female P. citri showed both H. zealandica and S. yirgalemense were able to complete their life cycles inside the host. Further bioassays illustrated a linear relationship between mealybug mortality and the concentration of nematodes applied, with the highest level of control using a concentration of 80 infective juveniles (IJs)/insect. As nematodes would be used as an above-ground application to control P. citri in citrus orchards, available water is a major limiting factor. Insecticidal activity proved to be dependent on the available surface moisture after nematode application. The water activity (a(w)) bioassay indicated that S. yirgalemense to be two times more tolerant to lower levels of free water, with a(w50)=0.96 and a(w90)=0.99, compared to H. zealandica with a(w50)=0.98 and a(w)90=1.0. After application, nematodes have a limited time frame in which to locate and infect hosts, as the level of available free water gradually decreases, as trees dry out. S. yirgalemense proved able to locate and infect P. citri quicker than H. zealandica. Nematode activity was not significantly affected when exposed to 15°C, 20°C and 25°C. IJs were able to infect P. citri at an exposure time as short as half an hour. Results also showed that the first 2-4h post application is the most decisive time for establishing successful infection of mealybugs. This is the first report on the potential use of nematodes for the control of P. citri.

  15. Analyses of Numerical Responses and Main Life Parameters for Determining the Suppression of Amblyseius cucumeris on Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yan-xuan; LIN Jian-zhen; JI Jie; CHEN Xie; KANG Yu-mei

    2005-01-01

    In this paper, the numeral response and main parameters of experimental population life table were analyzed for determiningthe suppressing ability of Amblyseius cucumeris on Panonychus citri. The result showed that: (1) Under 21-31 ℃ and 1-9 prey densities/leaf fragment condition, the prey consumptions ofA. cucumeris increased with the temperature or prey density; (2) In 1:3 predator-prey ratio treatment, the suppression ofP. citri (Ro=34.0053; T=19.4369; t=3.8204) was rather weak, it was enhanced as the ratio over 5:30, and the populations ofP. citri in these treatments can be fully controlled within 4-5 days; (3) Under 25±1℃, 80-85% RH and 15L: 9D illumination conditions, the net reproductive rate, mean generation duration and the time for population double increase ofP. citri (Ro=34.0053; T=19.4369; t=3.8204) were higher than those ofA. cucumeris (Ro=21.8750; T=16.8943; t=3.7954). While the intrinsic rate of increase and finite rate of increase of the former (rm=0.1814; λ= 1.1989) were lower than those of the latter (rm=0.1826; λ= 1.2004). These results indicated that A. cucumeris is a desirable bio-control agent to suppress P. citri at lower population stage in citrus orchard.

  16. Expression patterns of genes involved in the defense and stress response of Spiroplasma citri infected Madagascar Periwinkle Catharanthus roseus.

    Science.gov (United States)

    Nejat, Naghmeh; Vadamalai, Ganesan; Dickinson, Matthew

    2012-01-01

    Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of periwinkle. The response of plants to S. citri infection is very little known at the transcriptome level. In this study, quantitative real-time PCR (RT-qPCR) was used to investigate the expression levels of four selected genes involved in defense and stress responses in naturally and experimentally Spiroplasma citri infected periwinkles. Strictosidine β-glucosidase involved in terpenoid indole alkaloids (TIAs) biosynthesis pathway showed significant upregulation in experimentally and naturally infected periwinkles. The transcript level of extensin increased in leaves of periwinkles experimentally infected by S. citri in comparison to healthy ones. A similar level of heat shock protein 90 and metallothionein expression was observed in healthy, naturally and experimentally spiroplasma-diseased periwinkles. Overexpression of Strictosidine β-glucosidase demonstrates the potential utility of this gene as a host biomarker to increase the fidelity of S. citri detection and can also be used in breeding programs to develop stable disease-resistance varieties.

  17. Expression Patterns of Genes Involved in the Defense and Stress Response of Spiroplasma citri Infected Madagascar Periwinkle Catharanthus roseus

    Directory of Open Access Journals (Sweden)

    Naghmeh Nejat

    2012-02-01

    Full Text Available Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of periwinkle. The response of plants to S. citri infection is very little known at the transcriptome level. In this study, quantitative real-time PCR (RT-qPCR was used to investigate the expression levels of four selected genes involved in defense and stress responses in naturally and experimentally Spiroplasma citri infected periwinkles. Strictosidine β-glucosidase involved in terpenoid indole alkaloids (TIAs biosynthesis pathway showed significant upregulation in experimentally and naturally infected periwinkles. The transcript level of extensin increased in leaves of periwinkles experimentally infected by S. citri in comparison to healthy ones. A similar level of heat shock protein 90 and metallothionein expression was observed in healthy, naturally and experimentally spiroplasma-diseased periwinkles. Overexpression of Strictosidine β-glucosidase demonstrates the potential utility of this gene as a host biomarker to increase the fidelity of S. citri detection and can also be used in breeding programs to develop stable disease-resistance varieties.

  18. Establishment of EMA-PCR detection method of viable cells of Xanthomonas citri subsp.citri%柑橘溃疡病菌EMA-PCR快速活体检测技术的建立

    Institute of Scientific and Technical Information of China (English)

    熊书; 王中康; 卢小林; 殷幼平

    2013-01-01

    传统PCR方法不能诊断柑橘溃疡病菌(Xanthomonas citri subsp.citri Gabriel)的死活状态,往往导致假阳性检测结果.本研究将特异性核酸染料叠氮溴化乙锭(ethidium monoazide bromide,EMA)与PCR技术结合,旨在建立柑橘溃疡病活菌的快速检测技术.根据柑橘溃疡病菌独有的保守蛋白基因设计特异性引物扩增出278 bp的靶带,PCR反应的检测下限为25个细胞/25 μL或2.75 pg/25 μL.EMA-PCR结果表明:当卤钨灯曝光时间1 min,EMA终浓度为1.0 mg/L时,能有效抑制1.0×108 cfu/mL死菌的扩增;当EMA的浓度小于30 mg/L时,EMA对上述相同浓度活菌靶基因的扩增没有明显的抑制.EMA-PCR对死活混合菌的扩增表明,活菌数在6.875×101~6.875×105 cfu/PCR范围时,荧光强度与混合体系中活菌的对数值有线性关系.基于以上建立的EMA-PCR活体检测技术,对疑似带病柑橘材料进行检测,结果发现能降低柑橘溃疡病菌检测过程中的假阳性,有望为柑橘溃疡病的检疫检验提供更科学的技术手段.%Conventional PCR method can not distinguish live cells from dead cells of Xanthononas citri subsp.citri Gabriel,and false positives results can easily be obtained in detection process.Ethidium monoazide bromide (EMA) can selectively inhibit PCR amplification of DNA from dead cells.Therefore,in this study,EMA was combined with PCR to detect viable cells of X.citri subsp.citri.Firstly,primers (Xcc R/Xcc F) specific to X.citri subsp.citri were designed to amplify a 278 bp fragment.The minimum limit of detection was 25 cells/25 μL PCR volume or 2.75 pg/25 μ L PCR volume.The results of EMA-PCR showed that the optimized light exposure time was at least 1 min,allowing crosslinking of DNA by the EMA in dead cells and photolysing the free EMA in solution.The minimum amount of EMA to completely inhibit the PCR amplification of DNA derived from heat-killed cells was 1.0 mg/L.EMA less than 30 mg/L did not inhibit the PCR amplification

  19. Responses of the Asian citrus psyllid to volatiles emitted by the flushing shoots of its rutaceous host plants.

    Science.gov (United States)

    Patt, J M; Sétamou, M

    2010-04-01

    Diaphorina citri Kuwayama (Hemiptera: Psyllidae) carries Candidatus liberibacter spp., the putative causal agents of Huanglongbing. D. citri reproduces and develops only on the flushing shoots of its rutaceous host plants. Here we examined whether D. citri is attracted to host plant odors and a mixture of synthetic terpenes. Tests conducted in a vertically oriented Y-tube olfactometer showed that both males and females preferentially entered the Y-tube arm containing the odor from the young shoots of Murraya paniculata (L.) Jack and Citrus limon L. Burm. f. cultivar Eureka. Only males exhibited a preference for the odor of C. sinensis L., whereas the odor of C. x paradisi MacFadyen cultivar Rio Red was not attractive to both sexes. The volatiles emitted by young shoots of grapefruit cultivar Rio Red, Meyer lemon (Citrus x limon L. Burm.f.), and M. paniculata were analyzed by gas chromatograph-mass spectrometry. The samples were comprised of monoterpenes, monoterpene esters, and sesquiterpenes. The number of compounds present varied from 2 to 17, whereas the total amount of sample collected over 6 h ranged from 5.6 to 119.8 ng. The quantitatively dominant constituents were (E)-beta-ocimene, linalool, linalyl acetate, and beta-caryophyllene. The attractiveness of a mixture of synthetic terpenes, modeled on the volatiles collected from M. paniculata, was evaluated in screened cages in a no-choice test. At three observation intervals, significantly more individuals were trapped on white targets scented with the mixture than on unscented targets. These results indicate the feasibility of developing D. citri attractants patterned on actual host plant volatiles.

  20. Field Experiments of 13 Pesticides for Control Asian Citrus Psylla(Diaphorina citri Kuwayama)%吡蚜酮等13种农药防治柑桔木虱田间药效试验

    Institute of Scientific and Technical Information of China (English)

    邓明学; 唐美欣; 谭有龙; 陈贵峰; 唐明丽

    2009-01-01

    柑桔黄龙病是世界性的毁灭性病害,柑桔木虱是其唯一自然传播媒介,控制柑桔木虱是柑桔黄龙病综合防治最重要的技术措施.柑桔木虱成虫的高效传病性、近距离扩散性和快速传病性,决定了用来防治柑桔木虱成虫的农药要求速效性好、防效高(药后24h的防效>95%).由于受到试验地虫源和试验条件的限制,柑桔木虱的有效防治药剂筛选严重滞后.2007-2008年做了13种农药的田间小区药效试验和3种农药的大面积应用试验,结果表明:增效联苯菊酯EC 25 mg(a.i)/L、雕杀EC 110 mg(a.i)/L、铁骑EC 83 mg(a.i)/L对柑桔木虱成虫药后24 h的防效达100%;喹硫磷EC 250 mg(a.i)/L、杀扑磷EC200 mg(a.i)/L、稻丰散EC 333 mg(a.i)/L对柑桔木虱成虫药后24 h的防效>95%;防效在94.9%~85%的有丁硫克百威EC 200 mg(a.i)/L、霸中霸EC 200 mg(a.i)/L;吡虫啉WP 100 mg(a.i)/L、阿维·丁硫EC 90 mg(a.i)/L、甲基嘧啶磷500mg(a.i)/L的防效分别为92.3%~74.4%、79.1%、73.1;防效低于40%的药剂有吡蚜酮WP 250mg(a.i)/L、印楝素EC 6 mg(a.i)/L.

  1. Field Experiments of 15 Pesticides for Control Asian Citrus Psylla (Diaphorina citri Kuwayama)%20%哒虱威乳油等15种农药防治柑桔木虱田间药效试验

    Institute of Scientific and Technical Information of China (English)

    邓明学; 石晓雨

    2007-01-01

    控制柑桔木虱是柑桔黄龙病综合防治最重要的技术措施.柑桔木虱成虫的高效传病性、近距离扩散性、终生带毒性和快速传病性,决定了用来防治柑桔木虱成虫的农药要求速效性好、防效高(药后24 h的防效>95%).由于受到试验地虫源和试验条件的限制,柑桔木虱的有效防治药剂筛选严重滞后.2006-2007年做了15种农药的田间药效试验,结果表明:哒虱威EC 200mg(a.i)/L、丁硫克百威EC400mg(a.i)/L对柑桔木虱成虫药后24h的防效>95%;防效在94.9%~90%的有异丙威200mg(a.i)/L、133mg(a.i)/L、丁硫克百威EC 250mg(a.i)/L;噻嗪酮250mg(a.i)/L的防效只有30.8%;防效低于30%的药剂有灭多威200mg(a.i)/L、鱼藤酮150mg(a.i)/L、机油乳剂950mg(a.i)/L、阿维菌素20mg(a.i)/L和石硫合剂1波美度.机油乳剂与哒虱威混用无增效作用.

  2. Evaluation of efficacy of 18 strains of entomopathogenic nematodes (Rhabditida) against Planococcus citri (Risso, 1813) (Hemiptera: Pseudococcidae) under laboratory conditions.

    Science.gov (United States)

    Barbosa Negrisoli, Carla Ruth de Carvalho; Negrisoli Júnior, Aldomario Santo; Botton, Marcos; Garcia, Mauro Silveira; Bernardi, Daniel

    2013-07-01

    Planococcus citri (Risso, 1813) (Hemiptera: Pseudococcidae) is an important plant virus vector in grapevine crops in Brazil and other countries. The mealybug grows in roots and leaves of the grapes. Entomopathogenic nematodes (EPNs) are efficient control agents against insects associated to the soil and could be applied with the same equipment used for chemical insecticides. The aim of this study was to select effective EPNs for controlling P. citri females in laboratory conditions (25±1°C, UR 60±10%). We tested 17 native [Steinernema rarum (6 strains), Steinernema glaseri, Steinernema feltiae, Steinernema riobrave, Steinernema sp., Heterorhabditis bacteriophora (7 strains)] and only one exotic strain (Steinernema carpocapsae ALL). The bioassays were done on Petri dishes infested with females of P. citri, which were sprayed with EPNs juveniles. The strain with larger pathogenicity and virulence in laboratory was H. bacteriophora RS33 (from 69.0% to 92.2% of mortality), native of Rio Grande do Sul.

  3. Structural and Physiological Analyses of the Alkanesulphonate-Binding Protein (SsuA) of the Citrus Pathogen Xanthomonas citri

    Science.gov (United States)

    Tófoli de Araújo, Fabiano; Bolanos-Garcia, Victor M.; Pereira, Cristiane T.; Sanches, Mario; Oshiro, Elisa E.; Ferreira, Rita C. C.; Chigardze, Dimitri Y.; Barbosa, João Alexandre Gonçalves; de Souza Ferreira, Luís Carlos; Benedetti, Celso E.; Blundell, Tom L.; Balan, Andrea

    2013-01-01

    Background The uptake of sulphur-containing compounds plays a pivotal role in the physiology of bacteria that live in aerobic soils where organosulfur compounds such as sulphonates and sulphate esters represent more than 95% of the available sulphur. Until now, no information has been available on the uptake of sulphonates by bacterial plant pathogens, particularly those of the Xanthomonas genus, which encompasses several pathogenic species. In the present study, we characterised the alkanesulphonate uptake system (Ssu) of Xanthomonas axonopodis pv. citri 306 strain (X. citri), the etiological agent of citrus canker. Methodology/Principal Findings A single operon-like gene cluster (ssuEDACB) that encodes both the sulphur uptake system and enzymes involved in desulphurisation was detected in the genomes of X. citri and of the closely related species. We characterised X. citri SsuA protein, a periplasmic alkanesulphonate-binding protein that, together with SsuC and SsuB, defines the alkanesulphonate uptake system. The crystal structure of SsuA bound to MOPS, MES and HEPES, which is herein described for the first time, provides evidence for the importance of a conserved dipole in sulphate group coordination, identifies specific amino acids interacting with the sulphate group and shows the presence of a rather large binding pocket that explains the rather wide range of molecules recognised by the protein. Isolation of an isogenic ssuA-knockout derivative of the X. citri 306 strain showed that disruption of alkanesulphonate uptake affects both xanthan gum production and generation of canker lesions in sweet orange leaves. Conclusions/Significance The present study unravels unique structural and functional features of the X. citri SsuA protein and provides the first experimental evidence that an ABC uptake system affects the virulence of this phytopathogen. PMID:24282519

  4. Structural and physiological analyses of the alkanesulphonate-binding protein (SsuA of the citrus pathogen Xanthomonas citri.

    Directory of Open Access Journals (Sweden)

    Fabiano Tófoli de Araújo

    Full Text Available BACKGROUND: The uptake of sulphur-containing compounds plays a pivotal role in the physiology of bacteria that live in aerobic soils where organosulfur compounds such as sulphonates and sulphate esters represent more than 95% of the available sulphur. Until now, no information has been available on the uptake of sulphonates by bacterial plant pathogens, particularly those of the Xanthomonas genus, which encompasses several pathogenic species. In the present study, we characterised the alkanesulphonate uptake system (Ssu of Xanthomonas axonopodis pv. citri 306 strain (X. citri, the etiological agent of citrus canker. METHODOLOGY/PRINCIPAL FINDINGS: A single operon-like gene cluster (ssuEDACB that encodes both the sulphur uptake system and enzymes involved in desulphurisation was detected in the genomes of X. citri and of the closely related species. We characterised X. citri SsuA protein, a periplasmic alkanesulphonate-binding protein that, together with SsuC and SsuB, defines the alkanesulphonate uptake system. The crystal structure of SsuA bound to MOPS, MES and HEPES, which is herein described for the first time, provides evidence for the importance of a conserved dipole in sulphate group coordination, identifies specific amino acids interacting with the sulphate group and shows the presence of a rather large binding pocket that explains the rather wide range of molecules recognised by the protein. Isolation of an isogenic ssuA-knockout derivative of the X. citri 306 strain showed that disruption of alkanesulphonate uptake affects both xanthan gum production and generation of canker lesions in sweet orange leaves. CONCLUSIONS/SIGNIFICANCE: The present study unravels unique structural and functional features of the X. citri SsuA protein and provides the first experimental evidence that an ABC uptake system affects the virulence of this phytopathogen.

  5. Desenvolvimento de Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae em cafeeiros = Development of Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae in coffee trees

    Directory of Open Access Journals (Sweden)

    Lenira Viana Costa Santa-Cecília

    2009-01-01

    Full Text Available A cochonilha Planococcus citri (Risso, 1813 suga a seiva dos botões florais e frutos do cafeeiro, atacando as rosetas desde a floração até a colheita. Embora seja relatada há alguns anos na cafeicultura, são escassas as informações sobre o desenvolvimento dessacochonilha em cafeeiros. Assim, o presente trabalho teve por objetivos avaliar alguns aspectos biológicos da fase ninfal de P. citri em plantas de café. Ovos dessa cochonilha foram retirados de uma criação em laboratório, isolados em placas de Petri contendo discos foliares de Coffea arabica L., das cultivares Acaiá Cerrado, Mundo Novo e Catuaí Vermelho e de C. canephora Pierre & Froenher, cultivar Apoatã. As placas foram mantidas em câmara climatizada a 25 ± 1°C, 70 ± 10% de umidade relativa e 12h de fotofase. Constatou-se que a cultivar Catuaí Vermelho foi a que proporcionou maior duração do período ninfal das fêmeas, porém, não foram constatadas diferenças na mortalidade. Essa cochonilha se desenvolveu satisfatoriamente em todas as cultivares de café estudadas e os resultados não mostraram diferenças claras de susceptibilidade.The citrus mealybug Planococcus citri (Risso, 1813 feeds mainly on flowers and fruits of coffee plants from blooming until harvest. Little is known about its development in coffee although its occurrence is already known for several years. This work aims to study the nymph development of this mealybug in coffee plants.Eggs were isolated from a stock culture kept in laboratory and placed inside Petri dishes containing leaf sections in agar. The tested cultivars were Acaia Cerrado, Mundo Novo, Catuai Vermelho (Coffea arabica and Apoatã (C. canephora. Insects were kept in a climatizedchamber at 25ºC, 70 ± 10% humidity and 12-hour photophase. The longest development period in females was obtained in cultivar Catuai Vermelho. No differences in mortality were found among cultivars. Mealybugs developed in all cultivars and the results

  6. The abundant extrachromosomal DNA content of the Spiroplasma citri GII3-3X genome

    Directory of Open Access Journals (Sweden)

    Carrère Sébastien

    2008-04-01

    Full Text Available Abstract Background Spiroplama citri, the causal agent of citrus stubborn disease, is a bacterium of the class Mollicutes and is transmitted by phloem-feeding leafhopper vectors. In order to characterize candidate genes potentially involved in spiroplasma transmission and pathogenicity, the genome of S. citri strain GII3-3X is currently being deciphered. Results Assembling 20,000 sequencing reads generated seven circular contigs, none of which fit the 1.8 Mb chromosome map or carried chromosomal markers. These contigs correspond to seven plasmids: pSci1 to pSci6, with sizes ranging from 12.9 to 35.3 kbp and pSciA of 7.8 kbp. Plasmids pSci were detected as multiple copies in strain GII3-3X. Plasmid copy numbers of pSci1-6, as deduced from sequencing coverage, were estimated at 10 to 14 copies per spiroplasma cell, representing 1.6 Mb of extrachromosomal DNA. Genes encoding proteins of the TrsE-TraE, Mob, TraD-TraG, and Soj-ParA protein families were predicted in most of the pSci sequences, in addition to members of 14 protein families of unknown function. Plasmid pSci6 encodes protein P32, a marker of insect transmissibility. Plasmids pSci1-5 code for eight different S. citri adhesion-related proteins (ScARPs that are homologous to the previously described protein P89 and the S. kunkelii SkARP1. Conserved signal peptides and C-terminal transmembrane alpha helices were predicted in all ScARPs. The predicted surface-exposed N-terminal region possesses the following elements: (i 6 to 8 repeats of 39 to 42 amino acids each (sarpin repeats, (ii a central conserved region of 330 amino acids followed by (iii a more variable domain of about 110 amino acids. The C-terminus, predicted to be cytoplasmic, consists of a 27 amino acid stretch enriched in arginine and lysine (KR and an optional 23 amino acid stretch enriched in lysine, aspartate and glutamate (KDE. Plasmids pSci mainly present a linear increase of cumulative GC skew except in regions presenting

  7. Acción inhibitoria de una cepa de Zymomonas mobilis mobilis aislada de caña de azúcar sobre Xanthomonas citri subsp. citri, agente causal de la cancrosis de los cítricos Inhibition of Xanthomonas citri subsp. citri, causal agent of citrus canker, by a strain of Zymomonas mobilis mobilis isolated from sugarcane

    Directory of Open Access Journals (Sweden)

    María E. Romero

    2008-06-01

    Full Text Available Zymomonas mobilis mobilis (Zm produce factores antimicrobianos que actúan sobre un amplio espectro de microorganismos patógenos para el hombre, animales y plantas. Un problema importante a resolver en los tratamientos con antimicrobianos, es el desarrollo de resistencia a compuestos empleados actualmente, no siendo las bacterias fitopatógenas una excepción. En el presente trabajo se realizaron ensayos de antagonismo con células (pruebas de estrías cruzadas y sobrenadantes concentrados (Sc (por difusión en agar preparados a partir de cultivos de Zm (aislada de jugo de caña de azúcar producido en Tucumán, frente a la bacteria causal de la cancrosis: Xanthomonas citri subsp. citri. Se evaluaron aislamientos de Xcc sensibles (Xc y resistentes (Xcr a compuestos a base de cobre. Los resultados obtenidos mostraron que la bacteria testigo fue inhibida totalmente por las células de Zm, ejerciendo un efecto bactericida. En los ensayos de difusión en el agar se observó que tanto Xc, como Xcr fueron sensibles al Sc de Zm. Se sabe, por estudios anteriores, que los metabolitos de Zymomonas tienen un efecto deletéreo en la membrana celular de E. coli AB1133, inhibiéndose la respiración de la bacteria inmediatamente de agregado Sc (60 UA. En el presente trabajo se observó el mismo efecto, inhibición total de la respiración en Xc, luego del agregado del Sc (60 UA. Por lo observado, se deduce que el blanco de acción de los metabolitos antimicrobianos de Sc en Xc, sería el mismo que el de E. coli AB1133. Con los resultados obtenidos se considera de interés encarar el estudio de los compuestos de Zm para ser empleados en el control de enfermedades que afectan los cultivos de valor económico de la región, como es el caso de la cancrosis, como así también profundizar acerca de la acción de dichos metabolitos en la membrana de Xanthomonas citri subsp. citri.Zymomonas mobilis mobilis (Zm produces antimicrobial factors, which have an effect on

  8. An experimental test of rainfall as a control agent of Glycaspis brimblecombei Moore (Hemiptera, Psyllidae on seedlings of Eucalyptus camaldulensis Dehn (Myrtaceae

    Directory of Open Access Journals (Sweden)

    Karla N. Oliveira

    2012-03-01

    Full Text Available An experimental test of rainfall as a control agent of Glycaspis brimblecombei Moore (Hemiptera, Psyllidae on seedlings of Eucalyptus camaldulensis Dehn (Myrtaceae. Glycaspis brimblecombei is one the greatest threats to eucalyptus plantations in Brazil. The effects of rainfall to reduce the abundance of lerp of Glycaspis brimblecombei on experimentally infested seedlings of Eucalyptus camaldulensis were assessed. The number of lerps on the adaxial and abaxial surfaces of every leaf of 60 seedlings was recorded, before and after submission to the following treatments: "artificial rain", "leaf wetting" and control. A drastic reduction in lerp abundance per plant was observed after the treatments "leaf wetting" and artificial rain (F = 53.630; p Teste experimental da chuva como agente de controle de Glycaspis brimblecombei Moore (Hemiptera, Psyllidae em mudas de Eucalyptus camaldulensis Dehn (Myrtaceae. Glycaspis brimblecombei é uma das maiores ameaças das plantações de eucalipto do Brasil. Foram avaliados os efeitos da água na redução da abundância de conchas desse inseto em mudas de Eucalyptus camaldulensis infestadas experimentalmente. Foi quantificado o número de conchas nas superfícies adaxial e abaxial de todas as folhas de 60 mudas, antes e após a aplicação dos seguintes tratamentos: "chuva artificial", "molhamento das folhas" e controle. Foi observada uma drástica redução na abundância de conchas nos tratamentos "chuva artificial" e "molhamento das folhas" (F = 53,630; p < 0,001, o que não ocorreu para o tratamento controle ao longo do experimento (F = 1,450; p = 0,232. Ao final do experimento, a abundância de conchas foi significativamente menor no tratamento "chuva artificial" e "molhamento das folhas" do que no tratamento controle. Dessa forma, dois dias de chuva mostraram ser eficientes para diminuir mais que 50% da população de conchas, com quase 100% de eficiência após 5 dias de experimento. Nossos resultados

  9. Involvement of Three Esterase Genes from Panonychus citri (McGregor in Fenpropathrin Resistance

    Directory of Open Access Journals (Sweden)

    Xiao-Min Shen

    2016-08-01

    Full Text Available The citrus red mite, Panonychus citri (McGregor, is a major citrus pest with a worldwide distribution and an extensive record of pesticide resistance. However, the underlying molecular mechanism associated with fenpropathrin resistance in this species have not yet been reported. In this study, synergist triphenyl phosphate (TPP dramatically increased the toxicity of fenpropathrin, suggesting involvement of carboxylesterases (CarEs in the metabolic detoxification of this insecticide. The subsequent spatiotemporal expression pattern analysis of PcE1, PcE7 and PcE9 showed that three CarEs genes were all over-expressed after insecticide exposure and higher transcripts levels were observed in different field resistant strains of P. citri. Heterologous expression combined with 3-(4,5-dimethyl-thiazol-2-yl-2,5-diphenyltetra-zolium bromide (MTT cytotoxicity assay in Spodoptera frugiperda (Sf9 cells revealed that PcE1-, PcE7- or PcE9-expressing cells showed significantly higher cytoprotective capability than parental Sf9 cells against fenpropathrin, demonstrating that PcEs probably detoxify fenpropathrin. Moreover, gene silencing through the method of leaf-mediated dsRNA feeding followed by insecticide bioassay increased the mortalities of fenpropathrin-treated mites by 31% (PcE1, 27% (PcE7 and 22% (PcE9, respectively, after individual PcE gene dsRNA treatment. In conclusion, this study provides evidence that PcE1, PcE7 and PcE9 are functional genes mediated in fenpropathrin resistance in P. citri and enrich molecular understanding of CarEs during the resistance development of the mite.

  10. Involvement of Three Esterase Genes from Panonychus citri (McGregor) in Fenpropathrin Resistance.

    Science.gov (United States)

    Shen, Xiao-Min; Liao, Chong-Yu; Lu, Xue-Ping; Wang, Zhe; Wang, Jin-Jun; Dou, Wei

    2016-08-19

    The citrus red mite, Panonychus citri (McGregor), is a major citrus pest with a worldwide distribution and an extensive record of pesticide resistance. However, the underlying molecular mechanism associated with fenpropathrin resistance in this species have not yet been reported. In this study, synergist triphenyl phosphate (TPP) dramatically increased the toxicity of fenpropathrin, suggesting involvement of carboxylesterases (CarEs) in the metabolic detoxification of this insecticide. The subsequent spatiotemporal expression pattern analysis of PcE1, PcE7 and PcE9 showed that three CarEs genes were all over-expressed after insecticide exposure and higher transcripts levels were observed in different field resistant strains of P. citri. Heterologous expression combined with 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay in Spodoptera frugiperda (Sf9) cells revealed that PcE1-, PcE7- or PcE9-expressing cells showed significantly higher cytoprotective capability than parental Sf9 cells against fenpropathrin, demonstrating that PcEs probably detoxify fenpropathrin. Moreover, gene silencing through the method of leaf-mediated dsRNA feeding followed by insecticide bioassay increased the mortalities of fenpropathrin-treated mites by 31% (PcE1), 27% (PcE7) and 22% (PcE9), respectively, after individual PcE gene dsRNA treatment. In conclusion, this study provides evidence that PcE1, PcE7 and PcE9 are functional genes mediated in fenpropathrin resistance in P. citri and enrich molecular understanding of CarEs during the resistance development of the mite.

  11. The dual nature of trehalose in citrus canker disease: a virulence factor for Xanthomonas citri subsp. citri and a trigger for plant defence responses.

    Science.gov (United States)

    Piazza, Ainelén; Zimaro, Tamara; Garavaglia, Betiana S; Ficarra, Florencia A; Thomas, Ludivine; Marondedze, Claudius; Feil, Regina; Lunn, John E; Gehring, Chris; Ottado, Jorgelina; Gottig, Natalia

    2015-05-01

    Xanthomonas citri subsp. citri (Xcc) is a bacterial pathogen that causes citrus canker in susceptible Citrus spp. The Xcc genome contains genes encoding enzymes from three separate pathways of trehalose biosynthesis. Expression of genes encoding trehalose-6-phosphate synthase (otsA) and trehalose phosphatase (otsB) was highly induced during canker development, suggesting that the two-step pathway of trehalose biosynthesis via trehalose-6-phosphate has a function in pathogenesis. This pathway was eliminated from the bacterium by deletion of the otsA gene. The resulting XccΔotsA mutant produced less trehalose than the wild-type strain, was less resistant to salt and oxidative stresses, and was less able to colonize plant tissues. Gene expression and proteomic analyses of infected leaves showed that infection with XccΔotsA triggered only weak defence responses in the plant compared with infection with Xcc, and had less impact on the host plant's metabolism than the wild-type strain. These results suggested that trehalose of bacterial origin, synthesized via the otsA-otsB pathway, in Xcc, plays a role in modifying the host plant's metabolism to its own advantage but is also perceived by the plant as a sign of pathogen attack. Thus, trehalose biosynthesis has both positive and negative consequences for Xcc. On the one hand, it enables this bacterial pathogen to survive in the inhospitable environment of the leaf surface before infection and exploit the host plant's resources after infection, but on the other hand, it is a tell-tale sign of the pathogen's presence that triggers the plant to defend itself against infection. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  12. The dual nature of trehalose in citrus canker disease: a virulence factor for Xanthomonas citri subsp. citri and a trigger for plant defence responses

    KAUST Repository

    Piazza, A.

    2015-03-14

    Xanthomonas citri subsp. citri (Xcc) is a bacterial pathogen that causes citrus canker in susceptible Citrus spp. The Xcc genome contains genes encoding enzymes from three separate pathways of trehalose biosynthesis. Expression of genes encoding trehalose-6-phosphate synthase (otsA) and trehalose phosphatase (otsB) was highly induced during canker development, suggesting that the two-step pathway of trehalose biosynthesis via trehalose-6-phosphate has a function in pathogenesis. This pathway was eliminated from the bacterium by deletion of the otsA gene. The resulting XccΔotsA mutant produced less trehalose than the wild-type strain, was less resistant to salt and oxidative stresses, and was less able to colonize plant tissues. Gene expression and proteomic analyses of infected leaves showed that infection with XccΔotsA triggered only weak defence responses in the plant compared with infection with Xcc, and had less impact on the host plant\\'s metabolism than the wild-type strain. These results suggested that trehalose of bacterial origin, synthesized via the otsA-otsB pathway, in Xcc, plays a role in modifying the host plant\\'s metabolism to its own advantage but is also perceived by the plant as a sign of pathogen attack. Thus, trehalose biosynthesis has both positive and negative consequences for Xcc. On the one hand, it enables this bacterial pathogen to survive in the inhospitable environment of the leaf surface before infection and exploit the host plant\\'s resources after infection, but on the other hand, it is a tell-tale sign of the pathogen\\'s presence that triggers the plant to defend itself against infection. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  13. Responsiveness of different citrus genotypes to the Xanthomonas citri ssp. citri-derived pathogen-associated molecular pattern (PAMP) flg22 correlates with resistance to citrus canker.

    Science.gov (United States)

    Shi, Qingchun; Febres, Vicente J; Jones, Jeffrey B; Moore, Gloria A

    2015-06-01

    The bacterial agent of citrus canker disease (Xanthomonas citri ssp. citri, Xcc) has caused tremendous economic losses to the citrus industry around the world. Pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) is important to plant immunity. In this study, we compared the defence responses of citrus canker-resistant and citrus canker-susceptible genotypes to the Xcc-derived PAMP flg22 (Xflg22) by analysing the expression of 20 citrus defence-associated genes. We showed that, in the most resistant genotype, 'Nagami' kumquat, there was significant induction of several defence genes (EDS1, NDR1, PBS1, RAR1, SGT1, PAL1, NPR2 and NPR3) as early as 6 h and up to 72 h after Xflg22 treatment. At the other end of the spectrum, highly susceptible 'Duncan' grapefruit showed no induction of the same defence genes, even 120 h after treatment. Citrus genotypes with partial levels of resistance showed intermediate levels of transcriptional reprogramming that correlated with their resistance level. Xflg22 also triggered a rapid oxidative burst in all genotypes which was higher and accompanied by the induction of PTI marker genes (WRKY22 and GST1) only in the more resistant genotypes. Pretreatment with Xflg22 prior to Xcc inoculation inhibited bacterial growth in kumquat, but not in grapefruit. A flagellin-deficient Xcc strain (XccΔfliC) showed greater growth increase relative to wild-type Xcc in kumquat than in grapefruit. Taken together, our results indicate that Xflg22 initiates strong PTI in canker-resistant genotypes, but not in susceptible ones, and that a robust induction of PTI is an important component of citrus resistance to canker.

  14. Chemical composition and fungicidal activity of the essential oil of Thymus vulgaris against Alternaria citri

    Directory of Open Access Journals (Sweden)

    Erica A. Soto Mendívil

    2006-01-01

    Full Text Available Se analizó químicamente el aceite esencial de tomillo (Thymus vulgaris L. por Cromatografía de Gases/Espectroscopia de Masas y se evaluó su actividad fungicida. Los principales constituyentes fueron borneol (28.4%, timol (16.6%, carvacrol metil eter (9.6%, camfeno (6.9%, α-humulene (6.4% y carvacrol (5.0%. Las pruebas de actividad fungicida (in vitro indicaron que la concentración de 1000ppm del aceite esencial de tomillo fue efectivo para inhibir a Alternaria citri, cuando se adicionó al medio de cultivo agar papa dextrosa

  15. An experimental test of rainfall as a control agent of Glycaspis brimblecombei Moore (Hemiptera, Psyllidae on seedlings of Eucalyptus camaldulensis Dehn (Myrtaceae

    Directory of Open Access Journals (Sweden)

    Karla N. Oliveira

    2012-03-01

    Full Text Available An experimental test of rainfall as a control agent of Glycaspis brimblecombei Moore (Hemiptera, Psyllidae on seedlings of Eucalyptus camaldulensis Dehn (Myrtaceae. Glycaspis brimblecombei is one the greatest threats to eucalyptus plantations in Brazil. The effects of rainfall to reduce the abundance of lerp of Glycaspis brimblecombei on experimentally infested seedlings of Eucalyptus camaldulensis were assessed. The number of lerps on the adaxial and abaxial surfaces of every leaf of 60 seedlings was recorded, before and after submission to the following treatments: "artificial rain", "leaf wetting" and control. A drastic reduction in lerp abundance per plant was observed after the treatments "leaf wetting" and artificial rain (F = 53.630; p < 0.001, whereas lerp abundance remained roughly constant in the control treatment along the experiment (F = 1.450; p = 0.232. At the end of the experiment, lerp abundance was significantly lower in both the "artificial rain" and "leaf wetting" than in the control treatment. Two days of rainfall simulation were sufficient to decrease more than 50% of the lerp population, with almost 100% of effectiveness after 5 days of experiment. Our results indicate that lerp solubilization and mechanical removal by water are potential tools to the population regulation of G. brimblecombei on E. camaldulensis seedlings.

  16. Ultrastructure of Wax-Producing Structures on the Integument of the Melaleuca Psyllid Boreioglycaspis melaleucae (Hemiptera: Psyllidae), with Honeydew Excretion Behavior in Males and Females.

    Science.gov (United States)

    Ammar, El-Desouky; Hentz, Matthew; Hall, David G; Shatters, Robert G

    2015-01-01

    The melaleuca psyllid, Boreioglycaspis melaleucae (Hemiptera: Psyllidae), was introduced to Florida as a biological control agent against Melaleuca quinquenervia, an invasive evergreen tree that has invaded large areas of Florida Everglades. Colonies of B. melaleucae nymphs are normally covered by white waxy secretions, and nymphs of various instars produce long bundles of white waxy filaments extending laterally and posteriorly from their abdomen. Scanning electron microscopy of 'naturally waxed' and 'dewaxed' nymphs (cleaned from wax) revealed two types of wax pore plates located dorsally and laterally on the integument of posterior abdominal segments starting with the 4th segment. Type-1 wax pore plates, with raised rim, peripheral groove, slits and pits, produce long ribbons and filaments of waxy secretions that are wound together forming long wax bundles, whereas type-2 wax pore plates, with slits only, produce shorter wax curls. Additionally, in both nymphs and adult females, the circumanal ring contained ornate rows of wax pores that produce wax filaments covering their honeydew excretions. Video recordings with stereomicroscopy showed that adult females produce whitish honeydew balls, powerfully propelled away from their body, probably to get these sticky excretions away from their eggs and newly hatched nymphs. Adult males, however, produce clear droplets of honeydew immediately behind them, simply by bending the posterior end of the abdomen downward. The possible role(s) of waxy secretions by nymphs and adults of B. melaleucae in reducing contamination of their colonies with honeydew, among other possibilities, are discussed.

  17. Identification of differentially expressed genes of Xanthomonas axonopodis pv. citri by representational difference analysis of cDNA

    Directory of Open Access Journals (Sweden)

    Angela Mehta

    2005-03-01

    Full Text Available Xanthomonas axonopodis pv. citri is a phytopathogenic bacterium responsible for citrus canker, a serious disease which causes severe losses in citriculture around the world. In this study we report the differential expression of X. axonopodis pv. citri in response to specific treatments by using Representational Difference Analysis of cDNA (cDNA RDA. cDNAs from X. axonopodis pv. citri cultured in the presence of leaf extract of the host plant (Citrus sinensis, in vivo, as well as in the complex medium were hybridized against cDNA of the bacterium grown in the minimal medium. Sequencing of the difference products obtained after the second and third hybridizations revealed a total of 37 distinct genes identified by homology searches in the genome of X. axonopodis pv. citri. These genes were distributed in different functional categories, including genes that encode hypothetical proteins, genes involved in metabolism, cellular processes and pathogenicity, and mobile genetic elements. Most of these genes are likely related to growth and/or acquisition of nutrients in specific treatments whereas others might be important for the bacterium pathogenicity.

  18. Using mass spectrometry for identification of ABC transporters from Xanthomonas citri and mutants expressed in different growth conditions

    Energy Technology Data Exchange (ETDEWEB)

    Faria, J.N.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil); Paes Leme, A.F. [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil)

    2012-07-01

    Full text: Xanthomonas citri is a phytopathogenic bacterium that infects citrus plants causing significant losses for the economy. In our group, we have focused on the identification and characterization of ABC transport proteins of this bacterium, in order to determinate their function for growth in vitro and in vivo, during infection. ABC transporters represent one of the largest families of proteins, which transport since small molecules as ions up to oligopeptides and sugars. In prokaryotic cells many works have reported the ABC transport function in pathogenesis, resistance, biofilm formation, infectivity and DNA repair, but until our knowledge, there is no data related to these transporters and X. citri. So, In order to determinate which transporters are expressed in X. citri, we started a proteomic analysis based on mono and bi-dimensional gels associated to mass spectrometry analyses. After growing X. citri and two different mutants deleted for ssuA and nitA genes in LB and minimum media, cellular extracts were obtained and used for preparation of mono and bi-dimensional gels. Seven bands covering the expected mass of ABC transporter components (20 kDa to 50 kDa) in SDS-PAGE were cut off the gel, treated with trypsin and submitted to the MS for protein identification. The results of 2D gels were good enough and will serve as a standard for development of similar experiments in large scale. (author)

  19. Species limits in Diaporthe: molecular re-assessment of D. citri, D. cytosporella, D. foeniculina and D. rudis

    NARCIS (Netherlands)

    Udayanga, D.; Castlebury, L.A.; Rossman, A.Y.; Hyde, K.D.

    2014-01-01

    Species of Diaporthe are important plant pathogens of a wide range of hosts worldwide. In the present study the species causing melanose and stem end rot diseases of Citrus spp. are revised. Three species of Diaporthe occurring on Citrus are characterised, including D. citri, D. cytosporella and D.

  20. Identification and Characterization of Seven Glutathione S-Transferase Genes from Citrus Red Mite, Panonychus citri (McGregor

    Directory of Open Access Journals (Sweden)

    Chong-Yu Liao

    2013-12-01

    Full Text Available The citrus red mite, Panonychus citri (McGregor, is a global citrus pest, and has developed severe resistance to several types of acaricides. However, the molecular mechanisms of resistance in this mite remain unknown. In this study, seven full-length cDNAs encoding glutathione S-transferases (GSTs genes were identified and characterized in P. citri. The effects of pyridaben and fenpropathrin exposure on the expression of these genes were also investigated. Phylogenetic analysis revealed that the seven GSTs genes in P. citri cloned in this study belong to three different cytosolic classes, including four in mu, two in delta and one in zeta. Among these seven GSTs genes, the relative expression level of PcGSTm1 was significantly higher in adult than in the other life stages (egg, larvae and nymph. Compared with the control, the mRNA levels of the seven GST genes did not change significantly following exposure to pyridaben at LC10. However, RT-qPCR results showed that, when exposed to LC10 of fenpropathrin, six GSTs gene (PcGSTm1, PcGSTm3, PcGSTm4, PcGSTd1, PcGSTd2 and PcGSTz1 transcripts increased in a time-dependent manner. This is the first insight into the molecular characteristics of GSTs gene cDNAs in P. citri. The elevated GSTs gene transcripts following exposure to fenpropathrin might be one of the mechanisms involved in detoxification of this acaricide.

  1. Complete genome sequence of Spiroplasma citri strain R8-A2T, causal agent of stubborn disease in Citrus spp.

    Science.gov (United States)

    Spiroplasma citri is the causal agent of stubborn disease in Citrus spp., as well as the cause of diseases in numerous other plant genera. Here we report the nucleotide sequence of the 1,599,709 bp circular chromosome and two plasmids of strain R8-A2T. This information will facilitate comparative ...

  2. Characterization of the sulfate uptake and assimilation pathway from Xanthomonas citri - targets for bacterial growth inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Tambascia, C.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

    2012-07-01

    Full text: Microorganisms require sulfur for growth and obtain it either for inorganic sulfate or organosulfur compounds. ATP-Binding Cassete (SulT family) or major facilitator superfamily-type (SulP) transporters are responsible for the sulfate transport into the cell. In Xanthomonas citri, the phytopathogenic bacterium that causes the canker citrus disease, there are no reports related to the importance of these transporters during in vitro or in vivo infection. We identified in X. citri genome all the genes that belong to the well-characterized cys regulon from Escherichia coli and Salmonella typhimurium, which includes three ABC transporters and all the enzymes necessary for sulfate oxide reduction to sulfide and cysteine. Once these genes have been shown to be extremely important for bacteria growth and development in different environments, we chose the sbpcysWUA and cysDNCHIJG operons, which encodes the ABC inorganic sulfate ABC transporter and all the enzymes necessary for conversion of sulfate in cysteine, respectively. As a step for crystallization trials and resolution of their tridimensional structures, the referred genes were amplified and cloned into the cloning vector pGEM T-easy. In addition, using bioinformatics tools and molecular modeling we characterized all the protein functions as well as built tridimensional models of their structure for determination of the active sites. The importance of each protein is discussed aiming the discovery of a good target for development of inhibitors that could block the bacterium growth. (author)

  3. Development and reproduction of Panonychus citri (Prostigmata: Tetranychidae) on different species and varieties of citrus plants.

    Science.gov (United States)

    Zanardi, Odimar Zanuzo; Bordini, Gabriela Pavan; Franco, Aline Aparecida; de Morais, Matheus Rovere; Yamamoto, Pedro Takao

    2015-12-01

    The species and varieties of citrus plants that are currently grown can favor the population growth of the citrus red mite Panonychus citri (McGregor) (Prostigmata: Tetranychidae) and alter the pest management programs in citrus groves. In this study we evaluated, in the laboratory, the development and reproduction of P. citri and estimated its life table parameters when reared on four varieties of Citrus sinensis (L.) Osbeck (Valencia, Pera, Natal, and Hamlin), one variety of Citrus reticulata Blanco (Ponkan) and one variety of Citrus limon (L.) Burm. (Sicilian). The incubation period and egg viability were not affected by the host plant. However, the development and survival of the immature stage were significantly lower on Hamlin orange than on Valencia, Pera and Natal oranges, Ponkan mandarin and Sicilian lemon. The fecundity and oviposition period of females were lower on Hamlin orange than on the other hosts. Mites reared on Valencia orange and Sicilian lemon had a higher net reproductive rate (R 0 ), intrinsic growth rate (r) and finite rate of increase (λ), and a shorter interval between generations (T) than on Pera, Natal and Hamlin oranges and Ponkan mandarin. On the other hand, mites reared on Hamlin orange had the lowest R 0 , r and λ and the highest T among the hosts. Based on the results obtained we recommend that for Valencia orange and Sicilian lemon, the mite monitoring programs should be more intense to detect the initial infestation of pest, avoiding the damage in plants and the increase in production costs.

  4. A novel two-component response regulator links rpf with biofilm formation and virulence of Xanthomonas axonopodis pv. citri.

    Directory of Open Access Journals (Sweden)

    Tzu-Pi Huang

    Full Text Available Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a serious disease that impacts citrus production worldwide, and X. axonopodis pv. citri is listed as a quarantine pest in certain countries. Biofilm formation is important for the successful development of a pathogenic relationship between various bacteria and their host(s. To understand the mechanisms of biofilm formation by X. axonopodis pv. citri strain XW19, the strain was subjected to transposon mutagenesis. One mutant with a mutation in a two-component response regulator gene that was deficient in biofilm formation on a polystyrene microplate was selected for further study. The protein was designated as BfdR for biofilm formation defective regulator. BfdR from strain XW19 shares 100% amino acid sequence identity with XAC1284 of X. axonopodis pv. citri strain 306 and 30-100% identity with two-component response regulators in various pathogens and environmental microorganisms. The bfdR mutant strain exhibited significantly decreased biofilm formation on the leaf surfaces of Mexican lime compared with the wild type strain. The bfdR mutant was also compromised in its ability to cause canker lesions. The wild-type phenotype was restored by providing pbfdR in trans in the bfdR mutant. Our data indicated that BfdR did not regulate the production of virulence-related extracellular enzymes including amylase, lipase, protease, and lecithinase or the expression of hrpG, rfbC, and katE; however, BfdR controlled the expression of rpfF in XVM2 medium, which mimics cytoplasmic fluids in planta. In conclusion, biofilm formation on leaf surfaces of citrus is important for canker development in X. axonopodis pv. citri XW19. The process is controlled by the two-component response regulator BfdR via regulation of rpfF, which is required for the biosynthesis of a diffusible signal factor.

  5. A LOV protein modulates the physiological attributes of Xanthomonas axonopodis pv. citri relevant for host plant colonization.

    Directory of Open Access Journals (Sweden)

    Ivana Kraiselburd

    Full Text Available Recent studies have demonstrated that an appropriate light environment is required for the establishment of efficient vegetal resistance responses in several plant-pathogen interactions. The photoreceptors implicated in such responses are mainly those belonging to the phytochrome family. Data obtained from bacterial genome sequences revealed the presence of photosensory proteins of the BLUF (Blue Light sensing Using FAD, LOV (Light, Oxygen, Voltage and phytochrome families with no known functions. Xanthomonas axonopodis pv. citri is a Gram-negative bacterium responsible for citrus canker. The in silico analysis of the X. axonopodis pv. citri genome sequence revealed the presence of a gene encoding a putative LOV photoreceptor, in addition to two genes encoding BLUF proteins. This suggests that blue light sensing could play a role in X. axonopodis pv. citri physiology. We obtained the recombinant Xac-LOV protein by expression in Escherichia coli and performed a spectroscopic analysis of the purified protein, which demonstrated that it has a canonical LOV photochemistry. We also constructed a mutant strain of X. axonopodis pv. citri lacking the LOV protein and found that the loss of this protein altered bacterial motility, exopolysaccharide production and biofilm formation. Moreover, we observed that the adhesion of the mutant strain to abiotic and biotic surfaces was significantly diminished compared to the wild-type. Finally, inoculation of orange (Citrus sinensis leaves with the mutant strain of X. axonopodis pv. citri resulted in marked differences in the development of symptoms in plant tissues relative to the wild-type, suggesting a role for the Xac-LOV protein in the pathogenic process. Altogether, these results suggest the novel involvement of a photosensory system in the regulation of physiological attributes of a phytopathogenic bacterium. A functional blue light receptor in Xanthomonas spp. has been described for the first time, showing

  6. Signalering van citruswolluis (Planococcus citri) in de teelt van diverse potplanten : onderzoek binnen project 41203147 "Verbetering biologische bestrijding van wolluis in diverse potplanten”

    NARCIS (Netherlands)

    Boertjes, B.C.; Bruin, de J.

    2003-01-01

    In 2002 en 2003 is door PPO Glastuinbouw het project “Verbetering biologische bestrijding van wolluis in diverse potplanten” (project 41203147) uitgevoerd. Binnen dit project werd onder meer onderzoek gedaan naar methoden om citruswolluis (Planococcus citri) te signaleren.

  7. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves.

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    Full Text Available Xanthomonas citri subsp. citri (Xcc is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG, carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance.

  8. The gpsX gene encoding a glycosyltransferase is important for polysaccharide production and required for full virulence in Xanthomonas citri subsp. citri

    Directory of Open Access Journals (Sweden)

    Li Jinyun

    2012-03-01

    Full Text Available Abstract Background The Gram-negative bacterium Xanthomonas citri subsp. citri (Xac causes citrus canker, one of the most destructive diseases of citrus worldwide. In our previous work, a transposon mutant of Xac strain 306 with an insertion in the XAC3110 locus was isolated in a screening that aimed at identifying genes related to biofilm formation. The XAC3110 locus was named as bdp24 for biofilm-defective phenotype and the mutant was observed to be affected in extracellular polysaccharide (EPS and lipopolysaccharide (LPS biosynthesis and cell motility. In this study, we further characterized the bdp24 (XAC3110 gene (designated as gpsX using genetic complementation assays and expanded the knowledge about the function of the gpsX gene in Xac pathogenesis by investigating the roles of gpsX in EPS and LPS production, cell motility, biofilm formation on host leaves, stress tolerance, growth in planta, and host virulence of the citrus canker bacterium. Results The gpsX gene encodes a putative glycosyltransferase, which is highly conserved in the sequenced strains of Xanthomonas. Mutation of gpsX resulted in a significant reduction of the amount of EPS and loss of two LPS bands visualized on sodium dodecylsulphate- polyacrylamide gels. Biofilm assays revealed that the gpsX mutation affected biofilm formation by Xac on abiotic and biotic surfaces. The gpsX mutant showed delayed bacterial growth and caused reduced development of disease symptoms in susceptible citrus leaves. The gpsX mutant was more sensitive than the wild-type strain to various stresses, including the H2O2 oxidative stress. The mutant also showed attenuated ability in cell motility but not in flagellar formation. Quantitative reverse transcription-PCR assays indicated that mutation of gpsX did not affect the expression of virulence genes such as pthA in Xac strain 306. The affected phenotypes of the gpsX mutant could be complemented to wild-type levels by the intact gpsX gene

  9. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves.

    Science.gov (United States)

    Tondo, María Laura; Delprato, María Laura; Kraiselburd, Ivana; Fernández Zenoff, María Verónica; Farías, María Eugenia; Orellano, Elena G

    2016-01-01

    Xanthomonas citri subsp. citri (Xcc) is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS) at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV) radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG), carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance.

  10. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves

    Science.gov (United States)

    Tondo, María Laura; Delprato, María Laura; Kraiselburd, Ivana; Fernández Zenoff, María Verónica; Farías, María Eugenia; Orellano, Elena G.

    2016-01-01

    Xanthomonas citri subsp. citri (Xcc) is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS) at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV) radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG), carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance. PMID:26990197

  11. Overexpression of citrus polygalacturonase-inhibiting protein in citrus black rot pathogen Alternaria citri.

    Science.gov (United States)

    Katoh, Hiroshi; Nalumpang, Sarunya; Yamamoto, Hiroyuki; Akimitsu, Kazuya

    2007-05-01

    The rough lemon (Citrus jambhiri) gene encoding polygalacturonase-inhibiting protein (RlemPGIPA) was overexpressed in the pathogenic fungus Alternaria citri. The overexpression mutant AcOPI6 retained the ability to utilize pectin as a sole carbon source, and the overexpression of polygalacturonase-inhibiting protein did not have any effect on the growth of AcOPI6 in potato dextrose and pectin medium. The pathogenicity of AcOPI6 to cause a black rot symptom in citrus fruits was also unchanged. Polygalacturonase-inhibiting protein was secreted together with endopolygalacturonase into culture filtrates of AcOPI6, and oligogalacturonides were digested from polygalacturonic acid by both proteins in the culture filtrates. The reaction mixture containing oligogalacturonides possessed activity for induction of defense-related gene, RlemLOX, in rough lemon leaves.

  12. Chemotactic signal transduction and phosphate metabolism as adaptive strategies during citrus canker induction by Xanthomonas citri.

    Science.gov (United States)

    Moreira, Leandro Marcio; Facincani, Agda Paula; Ferreira, Cristiano Barbalho; Ferreira, Rafael Marine; Ferro, Maria Inês Tiraboshi; Gozzo, Fabio Cesar; de Oliveira, Julio Cezar Franco; Ferro, Jesus Aparecido; Soares, Márcia Regina

    2015-03-01

    The genome of Xanthomonas citri subsp. Citri strain 306 pathotype A (Xac) was completely sequenced more than 10 years; to date, few studies involving functional genomics Xac and its host compatible have been developed, specially related to adaptive events that allow the survival of Xac within the plant. Proteomic analysis of Xac showed that the processes of chemotactic signal transduction and phosphate metabolism are key adaptive strategies during the interaction of a pathogenic bacterium with its plant host. The results also indicate the importance of a group of proteins that may not be directly related to the classical virulence factors, but that are likely fundamental to the success of the initial stages of the infection, such as methyl-accepting chemotaxis protein (Mcp) and phosphate specific transport (Pst). Furthermore, the analysis of the mutant of the gene pstB which codifies to an ABC phosphate transporter subunit revealed a complete absence of citrus canker symptoms when inoculated in compatible hosts. We also conducted an in silico analysis which established the possible network of genes regulated by two-component systems PhoPQ and PhoBR (related to phosphate metabolism), and possible transcriptional factor binding site (TFBS) motifs of regulatory proteins PhoB and PhoP, detaching high degree of conservation of PhoB TFBS in 84 genes of Xac genome. This is the first time that chemotaxis signal transduction and phosphate metabolism were therefore indicated to be fundamental to the process of colonization of plant tissue during the induction of disease associated with Xanthomonas genus bacteria.

  13. Crystal Structure of the FAD-Containing Ferredoxin-NADP+ Reductase from the Plant Pathogen Xanthomonas axonopodis pv. citri

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    2013-01-01

    Full Text Available We have solved the structure of ferredoxin-NADP(H reductase, FPR, from the plant pathogen Xanthomonas axonopodis pv. citri, responsible for citrus canker, at a resolution of 1.5 Å. This structure reveals differences in the mobility of specific loops when compared to other FPRs, probably unrelated to the hydride transfer process, which contributes to explaining the structural and functional divergence between the subclass I FPRs. Interactions of the C-terminus of the enzyme with the phosphoadenosine of the cofactor FAD limit its mobility, thus affecting the entrance of nicotinamide into the active site. This structure opens the possibility of rationally designing drugs against the X. axonopodis pv. citri phytopathogen.

  14. De novo characterization of the Dialeurodes citri transcriptome: mining genes involved in stress resistance and simple sequence repeats (SSRs) discovery.

    Science.gov (United States)

    Chen, E-H; Wei, D-D; Shen, G-M; Yuan, G-R; Bai, P-P; Wang, J-J

    2014-02-01

    The citrus whitefly, Dialeurodes citri (Ashmead), is one of the three economically important whitefly species that infest citrus plants around the world; however, limited genetic research has been focused on D. citri, partly because of lack of genomic resources. In this study, we performed de novo assembly of a transcriptome using Illumina paired-end sequencing technology (Illumina Inc., San Diego, CA, USA). In total, 36,766 unigenes with a mean length of 497 bp were identified. Of these unigenes, we identified 17,788 matched known proteins in the National Center for Biotechnology Information database, as determined by Blast search, with 5731, 4850 and 14,441 unigenes assigned to clusters of orthologous groups (COG), gene ontology (GO), and SwissProt, respectively. In total, 7507 unigenes were assigned to 308 known pathways. In-depth analysis of the data showed that 117 unigenes were identified as potentially involved in the detoxification of xenobiotics and 67 heat shock protein (Hsp) genes were associated with environmental stress. In addition, these enzymes were searched against the GO and COG database, and the results showed that the three major detoxification enzymes and Hsps were classified into 18 and 3, 6, and 8 annotations, respectively. In addition, 149 simple sequence repeats were detected. The results facilitate the investigation of molecular resistance mechanisms to insecticides and environmental stress, and contribute to molecular marker development. The findings greatly improve our genetic understanding of D. citri, and lay the foundation for future functional genomics studies on this species.

  15. Sobre o condicionamento alimentar na cochonilha-branca, Planococcus citri (Risso (Hemiptera: pseudococcidae Regarding to host conditioning in citrus mealybug Planococcus citri (Risso (Hemiptera: pseudococcidae

    Directory of Open Access Journals (Sweden)

    Lenira Viana Costa Santa-Cecília

    2013-03-01

    Full Text Available Estudos do comportamento alimentar mediante a técnica de "Electrical Penetration Graphs" (EPG das cochonilhas-farinhentas (Pseudococcidae provenientes de um hospedeiro de criação alternativo têm mostrado que esses insetos não atingem ou demoram cerca de 9 horas para alcançar a fase floemática. Por outro lado, aqueles provenientes do hospedeiro-fonte atingem a fase floemática mais rapidamente e apresentam maior frequência de alimentação nos vasos crivados. Esses resultados indicam a presença do fenômeno de condicionamento alimentar, ainda não demonstrado em cochonilhas. Assim, o presente trabalho teve como objetivo determinar a existência desse fenômeno em Planococcus citri (Risso (Hemiptera: Pseudococcidae. Foram realizados testes de livre escolha, monitoramento eletrônico (EPG e estudos de alguns parâmetros biológicos. Em todos os experimentos, o cafeeiro (Coffea arabica L., os citros (Citrus sinensis L. e abóbora (Cucurbita maxima L. foram utilizados como substratos de criação (fonte da cochonilha, sendo os tratamentos constituídos pela combinação entre os hospedeiros-fonte e os hospedeiros receptores (café e citros. O teste de escolha entre cafeeiro e citros nas primeiras 72 horas mostrou que as cochonilhas criadas em cafeeiro apresentaram preferência pelo cafeeiro; aquelas originadas dos citros mostraram uma tendência, embora não significativa, em selecionar os citros em relação ao cafeeiro e aquelas criadas em abóbora não mostraram preferência por nenhum dos hospedeiros. Os estudos do comportamento alimentar mediante o monitoramento eletrônico (EPG mostraram que a fase floemática, considerada como a fase de aceitação do hospedeiro, foi mais frequente em cafeeiro, seja com cochonilhas oriundas deste substrato, seja de citros. Aqueles insetos mantidos em abóbora e transferidos para o cafeeiro ou citros apresentaram excepcionalmente ou não apresentaram nenhuma fase floemática, respectivamente. A

  16. The phloem-sap feeding mealybug (Ferrisia virgata carries 'Candidatus Liberibacter asiaticus' populations that do not cause disease in host plants.

    Directory of Open Access Journals (Sweden)

    Marco Pitino

    Full Text Available 'Candidatus Liberibacter asiaticus' (Las is the primary causal agent of huanglongbing (HLB, the most devastating disease of citrus worldwide. There are three known insect vectors of the HLB-associated bacteria, and all are members of the Hemiptera: Diaphorina citri (Psyllidae, Trioza erytreae (Triozidae, and Cacopsylla (Psylla citrisuga (Psyllidae. In this study, we found that another hemipteran, the striped mealybug Ferrisia virgata (Cockerell (Hemiptera: Pseudococcidae, was able to acquire and retain Las bacteria. The bacterial titers were positively correlated with the feeding acquisition time on Las-infected leaf discs, with a two-weeks feeding period resulting in Ct values ranging from 23.1 to 36.1 (8.24 × 10(7 to 1.07 × 10(4 Las cells per mealybug. We further discovered that the prophage/phage populations of Las in the mealybugs were different from those of Las in psyllids based on Las prophage-specific molecular markers: infected psyllids harbored the Las populations with prophage/phage FP1 and FP2, while infected mealybugs carried the Las populations with the iFP3 being the dominant prophage/phage. As in the psyllids, Las bacteria were shown to move through the insect gut wall to the salivary glands after being ingested by the mealybug based on a time-course quantitative polymerase chain reaction (qPCR assay of the dissected digestive systems. However, Las populations transmitted by the mealybugs did not cause disease in host plants. This is the first evidence of genetic difference among Las populations harbored by different insect vectors and difference among Las populations with respect to whether or not they cause disease in host plants.

  17. The Phloem-Sap Feeding Mealybug (Ferrisia virgata) Carries ‘Candidatus Liberibacter asiaticus’ Populations That Do Not Cause Disease in Host Plants

    Science.gov (United States)

    Pitino, Marco; Hoffman, Michele T.; Zhou, Lijuan; Hall, David G.; Stocks, Ian C.; Duan, Yongping

    2014-01-01

    ‘Candidatus Liberibacter asiaticus’ (Las) is the primary causal agent of huanglongbing (HLB), the most devastating disease of citrus worldwide. There are three known insect vectors of the HLB-associated bacteria, and all are members of the Hemiptera: Diaphorina citri (Psyllidae), Trioza erytreae (Triozidae), and Cacopsylla (Psylla) citrisuga (Psyllidae). In this study, we found that another hemipteran, the striped mealybug Ferrisia virgata (Cockerell) (Hemiptera: Pseudococcidae), was able to acquire and retain Las bacteria. The bacterial titers were positively correlated with the feeding acquisition time on Las-infected leaf discs, with a two-weeks feeding period resulting in Ct values ranging from 23.1 to 36.1 (8.24×107 to 1.07×104 Las cells per mealybug). We further discovered that the prophage/phage populations of Las in the mealybugs were different from those of Las in psyllids based on Las prophage-specific molecular markers: infected psyllids harbored the Las populations with prophage/phage FP1 and FP2, while infected mealybugs carried the Las populations with the iFP3 being the dominant prophage/phage. As in the psyllids, Las bacteria were shown to move through the insect gut wall to the salivary glands after being ingested by the mealybug based on a time-course quantitative polymerase chain reaction (qPCR) assay of the dissected digestive systems. However, Las populations transmitted by the mealybugs did not cause disease in host plants. This is the first evidence of genetic difference among Las populations harbored by different insect vectors and difference among Las populations with respect to whether or not they cause disease in host plants. PMID:24465578

  18. Polymerase chain reaction-restriction fragment length polymorphism method to distinguish three mealybug groups within the Planococcus citri-P. minor species complex (Hemiptera: Coccoidea: Pseudococcidae).

    Science.gov (United States)

    Rung, A; Miller, D R; Scheffer, S J

    2009-02-01

    The mealybug species Planococcus citri (Risso) and Planococcus minor (Maskell) (Hemiptera: Coccoidea: Pseudococcidae) have special significance to U.S. quarantine and U.S. agriculture. Commonly intercepted at U.S. ports-of-entry, they are difficult to identify based on morphological characters. This study presents a molecular method for distinguishing P. citri, P. minor, and a genetically distinct group that is morphologically identical to P. citri, from Hawaii. This method uses polymerase chain reaction (PCR) followed by restriction fragment polymorphism analysis (RFLP) using the restriction enzymes BspH1, BsmH1, and HpH1. The resulting band patterns can be visualized in a 2% agarose gel and are sufficient to differentiate between the three entities mentioned above. PCR-RFLP diagnostics can be used for all life stages and is cheaper and faster than DNA sequencing.

  19. Improved annotation of the insect vector of citrus greening disease: Biocuration by a diverse genomics community

    Science.gov (United States)

    The Asian citrus psyllid (Diaphorina citri Kuwayama) is the insect vector of the bacterium Candidatus Liberibacter asiaticus (CLas), the pathogen associated with citrus Huanglongbing (HLB, citrus greening). HLB threatens citrus production worldwide. Suppression or reduction of the insect vector usin...

  20. Annotation of the Asian citrus psyllid genome reveals a reduced innate immune system

    Science.gov (United States)

    Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuway...

  1. Fluctuación y distribución espacio-temporal de Tuthillia cognata (Hemiptera, Psyllidae y de Ocyptamus persimilis (Diptera, Syrphidae en el cultivo de camu-camu Myrciaria dubia (Myrtaceae en Ucayali, Perú Fluctuation and temporal-spatial distribution of Tuthillia cognata (Hemiptera, Psyllidae and Ocyptamus persimilis (Diptera, Syrphidae on camu-camu Myrciaria dubia (Myrtaceae in Ucayali, Peru

    Directory of Open Access Journals (Sweden)

    Diana Pérez

    2009-12-01

    Full Text Available Tuthillia cognata Hodkinson, Brown & Burckhardt, 1986 (Hemiptera, Psyllidae es una plaga importante en el cultivo de camu-camu, Myrciaria dubia H.B.K. Mc Vaugh (Myrtaceae en la Amazonía Peruana. El objetivo del presente estudio fue determinar la fluctuación y la distribución espacio-temporal de T. cognata y de su controlador biológico Ocyptamus persimilis (Curran, 1930 (Diptera, Syrphidae, entre enero a noviembre del 2004 en los caseríos San Juan y Padre Bernardo, Pucallpa, Ucayali, Perú. El número de ninfas y adultos de T. cognata fue mayor en la época lluviosa que en la seca, pero no en el número de huevos, ni en el número de colonias. Los huevos, ninfas y adultos prefirieron el tercio superior de la planta en comparación con el tercio medio e inferior en ambos caseríos. La excepción fue la presencia de huevos en el caserío San Juan que no mostró preferencias por ninguno de los tercios. No se observaron diferencias en el porcentaje de infestación por T. cognata entre ambos caseríos y entre la época seca y lluviosa. En O. persimilis, no se encontró diferencias en el número de huevos, larvas y pupas entre la época seca y lluviosa. Se observó que los huevos, larvas y pupas prefirieron el tercio superior de la planta en comparación con el tercio medio e inferior en ambos caseríos. Ambas especies, T. cognata y O. persimilis presentaron un patrón de distribución espacial - temporal agregado. Se encontró una relación directa entre el número de colonias de T. cognata y O. persimilis. Sin embargo, el número de huevos, ninfas y adultos de T. cognata no se encontraron correlacionados con el número de huevos, larvas y pupas de O. persimilis.Tuthillia cognata Hodkinson, Brown & Burckhardt, 1986 (Hemiptera, Psyllidae, is a main pest on camu-camu culture Myrciaria dubia H.B.K. Mc Vaugh (Myrtaceae in the Peruvian Amazon. The aim of the current research was to evaluate the fluctuation and spatial and temporal distribution of T

  2. Costs and benefits of insecticide and foliar nutrient applications to huanglongbing-infected citrus trees.

    Science.gov (United States)

    Tansey, James A; Vanaclocha, Pilar; Monzo, Cesar; Jones, Moneen; Stansly, Philip A

    2017-05-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), vectors Candidatus Liberibacter asiaticus, which causes huanglongbing (HLB). In Florida, HLB incidence is approaching 100% statewide. Yields have decreased and production costs have increased since 2005. Despite this, some growers are maintaining a level of production and attribute this in part to aggressive psyllid control and foliar nutrition sprays. However, the value of these practices is debated. A replicated field study was initiated in 2008 in a commercial block of 'Valencia' sweet orange trees to evaluate individual and combined effects of foliar nutrition and ACP control. Results from 2012-2016 are presented. Insecticides consistently reduced ACP populations. However, neither insecticide nor nutrition applications significantly influenced HLB incidence or PCR copy number in mature trees. In reset trees, infection continued to build and reached 100% in all treatments. Greatest yields (kg fruit ha(-1) ) and production (kg solids ha(-1) ) were obtained from trees receiving both insecticides and foliar nutrition. All treatments resulted in production and financial gains relative to controls. However, material and application costs associated with the nutrition component offset these gains, resulting in lesser benefits than insecticides applied alone. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  3. The effect of Leptomastix dactylopii parasitism and venom injection on host Planococcus citri

    Directory of Open Access Journals (Sweden)

    D Battaglia

    2014-10-01

    Full Text Available One of the major alterations observed in mealybug Planococcus citri parasitized by Leptomastix dactylopii is a strong reduction of laid eggs, which is evident soon after parasitization. Venom injection in unparasitized hosts determines a drastic reduction of fecundity indicating that this female secretion injected at the oviposition plays a key-role in host regulation. In order to assess the impact of parasitism and venom injection on host reproductive tissues, ovaries were dissected at different time intervals after these treatments and observed by light and transmission electron microscopy. The developing eggs showed clear symptoms of degeneration, already half an hour after parasitization or venom injection. Heat and protease treatments of venom nearly suppressed its effects on host reproduction, indicating that proteins are likely responsible for the observed alterations. The electrophoretic profile of venom proteins covers a wide range of molecular masses between 15 to 200 kDa but five major bands having a molecular mass of about 27, 30, 40, 90 and 120 kDa respectively were more evident. Moreover, to establish any parasitoid preference in host selection, among the adult female mealybugs at different stages of maturation and a possible relation with fecundity reduction in the host, the parasitoid behavior was observed.

  4. The GC/MS Analysis of Volatile Components Extracted by Different Methods from Exocarpium Citri Grandis

    Directory of Open Access Journals (Sweden)

    Zhisheng Xie

    2013-01-01

    Full Text Available Volatile components from Exocarpium Citri Grandis (ECG were, respectively, extracted by three methods, that is, steam distillation (SD, headspace solid-phase microextraction (HS-SPME, and solvent extraction (SE. A total of 81 compounds were identified by gas chromatography-mass spectrometry including 77 (SD, 56 (HS-SPME, and 48 (SE compounds, respectively. Despite of the extraction method, terpenes (39.98~57.81% were the main volatile components of ECG, mainly germacrene-D, limonene, 2,6,8,10,14-hexadecapentaene, 2,6,11,15-tetramethyl-, (E,E,E-, and trans-caryophyllene. Comparison was made among the three methods in terms of extraction profile and property. SD relatively gave an entire profile of volatile in ECG by long-time extraction; SE enabled the analysis of low volatility and high molecular weight compounds but lost some volatiles components; HS-SPME generated satisfactory extraction efficiency and gave similar results to those of SD at analytical level when consuming less sample amount, shorter extraction time, and simpler procedure. Although SD and SE were treated as traditionally preparative extractive techniques for volatiles in both small batches and large scale, HS-SPME coupled with GC/MS could be useful and appropriative for the rapid extraction and qualitative analysis of volatile components from medicinal plants at analytical level.

  5. Identification of anti-asthmatic compounds in Pericarpium citri reticulatae and evaluation of their synergistic effects

    Institute of Scientific and Technical Information of China (English)

    Qian SHI; Ze LIU; Yang YANG; Peng GENG; Yuan-yuan ZHU; Qi ZHANG; Fang BAI; Gang BAI

    2009-01-01

    Aim: To investigate the anti-asthmatic mechanisms of the traditional Chinese medicine Pericarpium citri reticulatae (PCR).Methods: The alkaloid section (AS) of PCR was extracted using an ion exchange resin, separated, and purified into different fractions by semi-preparative HPLC. These fractions were screened for beta2-adrenergic receptor (β2AR) agonistic activity using rat β2AR-transfected CHO-CRE-EGFP cells. AS and its isolated components were characterized by ultra-performance liquid chromatography/quadrupole time-of-flight MS (UPLC/Q-Tof MS) and were evaluated for their spasmolytic and antitussive activities both in vitro and in vivo in a guinea pig model.Results: We demonstrated that the AS component responsible for activating β2AR signaling was synephrine. Both AS and synephrine showed significant spasmolytic effects on acetylcholine chloride (ACh)-induced contractions in isolated guinea pig trachea, and they protected against histamine-induced experimental asthma by prolonging the latent period. We further identified stachydrine as the antitussive component that could significantly reduce citric acid-induced coughing. The combination of these two bioactive compounds had a more potent spasmolytic activity in comparison with the single use of synephrine or stachydrine.Conclusion: We conclude that synephrine and stachydrine are the key components of AS that mediate asthma relief due to their synergism when used in combination.

  6. Genomic insights into the evolutionary origin of Xanthomonas axonopodis pv. citri and its ecological relatives.

    Science.gov (United States)

    Midha, Samriti; Patil, Prabhu B

    2014-10-01

    Xanthomonas axonopodis pv. citri (Xac) is the causal agent of citrus bacterial canker (CBC) and is a serious problem worldwide. Like CBC, several important diseases in other fruits, such as mango, pomegranate, and grape, are also caused by Xanthomonas pathovars that display remarkable specificity toward their hosts. While citrus and mango diseases were documented more than 100 years ago, the pomegranate and grape diseases have been known only since the 1950s and 1970s, respectively. Interestingly, diseases caused by all these pathovars were noted first in India. Our genome-based phylogenetic studies suggest that these diverse pathogens belong to a single species and these pathovars may be just a group of rapidly evolving strains. Furthermore, the recently reported pathovars, such as those infecting grape and pomegranate, form independent clonal lineages, while the citrus and mango pathovars that have been known for a long time form one clonal lineage. Such an understanding of their phylogenomic relationship has further allowed us to understand major and unique variations in the lineages that give rise to these pathovars. Whole-genome sequencing studies including ecological relatives from their putative country of origin has allowed us to understand the evolutionary history of Xac and other pathovars that infect fruits.

  7. Pathogenic Interactions Between Xanthomonas axonopodis pv. citri and Cultivars of Pummelo (Citrus grandis).

    Science.gov (United States)

    Shiotani, H; Ozaki, K; Tsuyumu, S

    2000-12-01

    ABSTRACT The aggressiveness of strains of Xanthomonas axonopodis pv. citri on seven Citrus species, including Citrus sinensis (navel orange), C. paradisi (grapefruit), C. unshiu (Satsuma mandarin), C. junos (Yuzu), C. aurantifolia ('Mexican' lime), C. tachibana (Tachibana), and C. grandis (pummelo: 'Otachibana', 'Banpeiyu', and 'Anseikan'), were assessed by comparing lesion expansion and growth in planta, using a prick inoculation method. The existence of two groups distinct in aggressiveness was demonstrated on the pummelo cultivars, whereas the remaining species tested were uniformly susceptible. The two groups of strains were distinct in lesion expansion and growth in planta; however, both caused canker lesions on the 'Otachibana' pummelo. The sensitivity of the bacterial strains to phages Cp1 and Cp2 was associated with differences in aggressiveness. Namely, all the strains sensitive to Cp2 but resistant to Cp1 were aggressive to 'Otachibana', whereas all the strains sensitive to Cp1 but resistant to Cp2 were weakly aggressive. When a repetitive sequence-based polymerase chain reaction amplification was carried out by enterobacterial repetitive intergeneric consensus (ERIC) sequences (ERIC1R and ERIC2) as the primers, these two groups were also distinguishable by the presence or absence of a 1.8-kb DNA fragment among otherwise identical fragments. The 1.8-kb fragment was amplified only from the strains aggressive to C. grandis.

  8. Remote Sensing of Low and Mid-Latitude Ionospheric Disturbances During Solar Minimum Using CITRIS and CERTO Measurements of TEC and Radio Scintillation

    Science.gov (United States)

    Siefring, C. L.; Bernhardt, P. A.

    2010-12-01

    Unique data on ionospheric plasma disturbances from the Naval Research Laboratory CITRIS (Scintillation and TEC Receiver in Space) instrument will be presented. CITRIS is a multi-band receiver that recorded TEC (Total Electron Content) and radio scintillations from Low-Earth Orbit (LEO) on STPSat1. The 555+/5 km altitude 35° inclination orbit covers low and mid-latitudes. The measurements require propagation from a transmitter to a receiver through the F-region plasma. CITRIS used both 1) satellite beacons in LEO, such as the NRL CERTO (Coherent Electromagnetic Radio TOmography) three-frequency beacons transmitting at 150/400/1067 MHz and 2) the French global network of ground-based DORIS (Doppler Orbitography and Radiopositioning Integrated by Satellite) beacons transmitting at 401.25 and 2036.25 MHz. CITRIS was operated in a complementary fashion with the C/NOFS satellite during most of its first year of operations; C/NOFS carries CERTO beacon along with in-situ diagnostics. CITRIS and ground receivers can simultaneously measure TEC and scintillations on different paths using CERTO on C/NOFS. When C/NOFS is not in view, CITRIS makes measurements from DORIS beacons and other LEO satellites. Because of the orbits CITRIS will always make measurements at the same longitude within 48 min of C/NOFS. The ability to look at multiple paths is unique and useful for studying the spatial extent and time duration of disturbances. The combination of TEC and scintillation measurements provides information on a range of scale-sizes from >1 km to about 100 m. The joint data set on plasma structures at low-latitudes is a focus of our presentation, with the addition of comparisons to CITRIS data taken at mid-latitude. Several types of irregularities have been studied including Spread-F and the newly discovered dawn-side depletions. The data covers large portions of the Earth (including the Pacific, African and South American sectors) during an unusually quite portion of the most

  9. Characterization of bacteriophages Cp1 and Cp2, the strain-typing agents for Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Ahmad, Abdelmonim Ali; Ogawa, Megumi; Kawasaki, Takeru; Fujie, Makoto; Yamada, Takashi

    2014-01-01

    The strains of Xanthomonas axonopodis pv. citri, the causative agent of citrus canker, are historically classified based on bacteriophage (phage) sensitivity. Nearly all X. axonopodis pv. citri strains isolated from different regions in Japan are lysed by either phage Cp1 or Cp2; Cp1-sensitive (Cp1(s)) strains have been observed to be resistant to Cp2 (Cp2(r)) and vice versa. In this study, genomic and molecular characterization was performed for the typing agents Cp1 and Cp2. Morphologically, Cp1 belongs to the Siphoviridae. Genomic analysis revealed that its genome comprises 43,870-bp double-stranded DNA (dsDNA), with 10-bp 3'-extruding cohesive ends, and contains 48 open reading frames. The genomic organization was similar to that of Xanthomonas phage phiL7, but it lacked a group I intron in the DNA polymerase gene. Cp2 resembles morphologically Escherichia coli T7-like phages of Podoviridae. The 42,963-bp linear dsDNA genome of Cp2 contained terminal repeats. The Cp2 genomic sequence has 40 open reading frames, many of which did not show detectable homologs in the current databases. By proteomic analysis, a gene cluster encoding structural proteins corresponding to the class III module of T7-like phages was identified on the Cp2 genome. Therefore, Cp1 and Cp2 were found to belong to completely different virus groups. In addition, we found that Cp1 and Cp2 use different molecules on the host cell surface as phage receptors and that host selection of X. axonopodis pv. citri strains by Cp1 and Cp2 is not determined at the initial stage by binding to receptors.

  10. 60Co-γ Irradiation Affects the Enzymatic Antioxidant System of the Citrus Red Mite Panonychus citri (Acari: Tetranychidae

    Directory of Open Access Journals (Sweden)

    Ke Zhang

    2014-05-01

    Full Text Available Radio-(60Co, which emits γ rays, has been used worldwide in pest control. The aim of the present study was to analyze the effect of effective-low-power 60Co-γ irradiation on the enzymatic antioxidant system of the citrus red mite Panonychus citri. One day old female adults were exposed to 0.4 kGy 60Co-γ irradiation and on the, 6th h, 1st day, 2nd day, and 5th day post treatment, the mites were euthanized for biochemical analysis. The activity of superoxide dismutase (SOD, catalase (CAT, peroxidase (POD, phenoloxidase (PO and acetylocholinesterase (AchE were investigated. POD and CAT activities, as well as SOD were higher in the irradiated mites. We found that exposure to 60Co-γ radiation resulted in increased activities of SOD, CAT, POD and decreased AchE activity. When the recovery time lasted till the 5th day, the activities of POD and PO were significantly lower than the control, whereas the SOD, CAT and AchE activities returned to control levels. Cells possess protein repair pathways to rescue oxidized proteins and restore their functions, but if these repair processes fail, oxidized proteins may become cytotoxic. Our results confirm the hypothesis that low dosages of 60Co-γ irradiation increase the level of oxidative stress in P. citri adults in a short time, causing meanwhile damage and sterility. The results of this study also indicate that stress caused by exposure to irradiation could inhibit the cholinergic system in P. citri.

  11. Multiplexed lateral flow microarray assay for detection of citrus pathogens Xylella fastidiosa and Xanthomonas axonopodis pv citri

    Energy Technology Data Exchange (ETDEWEB)

    Cary; R. Bruce (Santa Fe, NM); Stubben, Christopher J. (Los Alamos, NM)

    2011-03-22

    The invention provides highly sensitive and specific assays for the major citrus pathogens Xylella fastidiosa and Xanthomonas axonopodis, including a field deployable multiplexed assay capable of rapidly assaying for both pathogens simultaneously. The assays are directed at particular gene targets derived from pathogenic strains that specifically cause the major citrus diseases of citrus variegated chlorosis (Xylella fastidiosa 9a5c) and citrus canker (Xanthomonas axonopodis pv citri). The citrus pathogen assays of the invention offer femtomole sensitivity, excellent linear dynamic range, and rapid and specific detection.

  12. Structural-functional characterization and physiological significance of ferredoxin-NADP reductase from Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Tondo, María Laura; Musumeci, Matías A; Delprato, María Laura; Ceccarelli, Eduardo A; Orellano, Elena G

    2011-01-01

    Xanthomonas axonopodis pv. citri is a phytopathogen bacterium that causes severe citrus canker disease. Similar to other phytopathogens, after infection by this bacterium, plants trigger a defense mechanism that produces reactive oxygen species. Ferredoxin-NADP(+) reductases (FNRs) are redox flavoenzymes that participate in several metabolic functions, including the response to reactive oxygen species. Xanthomonas axonopodis pv. citri has a gene (fpr) that encodes for a FNR (Xac-FNR) that belongs to the subclass I bacterial FNRs. The aim of this work was to search for the physiological role of this enzyme and to characterize its structural and functional properties. The functionality of Xac-FNR was tested by cross-complementation of a FNR knockout Escherichia coli strain, which exhibit high susceptibility to agents that produce an abnormal accumulation of (•)O(2)(-). Xac-FNR was able to substitute for the FNR in E. coli in its antioxidant role. The expression of fpr in X. axonopodis pv. citri was assessed using semiquantitative RT-PCR and Western blot analysis. A 2.2-fold induction was observed in the presence of the superoxide-generating agents methyl viologen and 2,3-dimethoxy-1,4-naphthoquinone. Structural and functional studies showed that Xac-FNR displayed different functional features from other subclass I bacterial FNRs. Our analyses suggest that these differences may be due to the unusual carboxy-terminal region. We propose a further classification of subclass I bacterial FNRs, which is useful to determine the nature of their ferredoxin redox partners. Using sequence analysis, we identified a ferredoxin (XAC1762) as a potential substrate of Xac-FNR. The purified ferredoxin protein displayed the typical broad UV-visible spectrum of [4Fe-4S] clusters and was able to function as substrate of Xac-FNR in the cytochrome c reductase activity. Our results suggest that Xac-FNR is involved in the oxidative stress response of Xanthomonas axonopodis pv. citri and

  13. Structural-functional characterization and physiological significance of ferredoxin-NADP reductase from Xanthomonas axonopodis pv. citri.

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    Full Text Available Xanthomonas axonopodis pv. citri is a phytopathogen bacterium that causes severe citrus canker disease. Similar to other phytopathogens, after infection by this bacterium, plants trigger a defense mechanism that produces reactive oxygen species. Ferredoxin-NADP(+ reductases (FNRs are redox flavoenzymes that participate in several metabolic functions, including the response to reactive oxygen species. Xanthomonas axonopodis pv. citri has a gene (fpr that encodes for a FNR (Xac-FNR that belongs to the subclass I bacterial FNRs. The aim of this work was to search for the physiological role of this enzyme and to characterize its structural and functional properties. The functionality of Xac-FNR was tested by cross-complementation of a FNR knockout Escherichia coli strain, which exhibit high susceptibility to agents that produce an abnormal accumulation of (•O(2(-. Xac-FNR was able to substitute for the FNR in E. coli in its antioxidant role. The expression of fpr in X. axonopodis pv. citri was assessed using semiquantitative RT-PCR and Western blot analysis. A 2.2-fold induction was observed in the presence of the superoxide-generating agents methyl viologen and 2,3-dimethoxy-1,4-naphthoquinone. Structural and functional studies showed that Xac-FNR displayed different functional features from other subclass I bacterial FNRs. Our analyses suggest that these differences may be due to the unusual carboxy-terminal region. We propose a further classification of subclass I bacterial FNRs, which is useful to determine the nature of their ferredoxin redox partners. Using sequence analysis, we identified a ferredoxin (XAC1762 as a potential substrate of Xac-FNR. The purified ferredoxin protein displayed the typical broad UV-visible spectrum of [4Fe-4S] clusters and was able to function as substrate of Xac-FNR in the cytochrome c reductase activity. Our results suggest that Xac-FNR is involved in the oxidative stress response of Xanthomonas axonopodis pv

  14. Determination of Alternaria Brown Spot (Alternaria alternata f.sp. citri) Disease on Minneola Tangelo in Çukurova Region

    OpenAIRE

    ERKILIÇ, Ali; CANIHOŞ, Yeter; BİÇİCİ, Mehmet

    1998-01-01

    In this study, the incidence and severity of Alternaria brown spot disease (Alternaria alternata f.sp. citri) in the Çukurova region were determined. Thirty-five Minneola tangelo orchards, with a total area of 2497 da, in 24 different locations were surveyed for this purpose. Leaf infections in all orchards were observed in the spring flushing period. Disease severity was found to be 0-10 % in 10 orchards, 11-20 % in 10 orchards, 21-30 % in 7 orchards, 31-40 % in 5 orchards, 42.5 % in 1 orch...

  15. Sobre o condicionamento alimentar na cochonilha-branca, Planococcus citri (Risso (Hemiptera: pseudococcidae

    Directory of Open Access Journals (Sweden)

    Lenira Viana Costa Santa-Cecília

    2013-03-01

    Full Text Available Estudos do comportamento alimentar mediante a técnica de "Electrical Penetration Graphs" (EPG das cochonilhas-farinhentas (Pseudococcidae provenientes de um hospedeiro de criação alternativo têm mostrado que esses insetos não atingem ou demoram cerca de 9 horas para alcançar a fase floemática. Por outro lado, aqueles provenientes do hospedeiro-fonte atingem a fase floemática mais rapidamente e apresentam maior frequência de alimentação nos vasos crivados. Esses resultados indicam a presença do fenômeno de condicionamento alimentar, ainda não demonstrado em cochonilhas. Assim, o presente trabalho teve como objetivo determinar a existência desse fenômeno em Planococcus citri (Risso (Hemiptera: Pseudococcidae. Foram realizados testes de livre escolha, monitoramento eletrônico (EPG e estudos de alguns parâmetros biológicos. Em todos os experimentos, o cafeeiro (Coffea arabica L., os citros (Citrus sinensis L. e abóbora (Cucurbita maxima L. foram utilizados como substratos de criação (fonte da cochonilha, sendo os tratamentos constituídos pela combinação entre os hospedeiros-fonte e os hospedeiros receptores (café e citros. O teste de escolha entre cafeeiro e citros nas primeiras 72 horas mostrou que as cochonilhas criadas em cafeeiro apresentaram preferência pelo cafeeiro; aquelas originadas dos citros mostraram uma tendência, embora não significativa, em selecionar os citros em relação ao cafeeiro e aquelas criadas em abóbora não mostraram preferência por nenhum dos hospedeiros. Os estudos do comportamento alimentar mediante o monitoramento eletrônico (EPG mostraram que a fase floemática, considerada como a fase de aceitação do hospedeiro, foi mais frequente em cafeeiro, seja com cochonilhas oriundas deste substrato, seja de citros. Aqueles insetos mantidos em abóbora e transferidos para o cafeeiro ou citros apresentaram excepcionalmente ou não apresentaram nenhuma fase floemática, respectivamente. A

  16. New occupational allergen in citrus farmers: citrus red mite (Panonychus citri).

    Science.gov (United States)

    Kim, Y K; Son, J W; Kim, H Y; Park, H S; Lee, M H; Cho, S H; Min, K U; Kim, Y Y

    1999-02-01

    There have been several reports of occupational allergy to spider mites (Tetranychidae), but no published report has described citrus red mite (CRM, Panonychus citri)-induced occupational asthma confirmed by specific bronchial challenge. The purpose of this study was to evaluate clinical and immunologic characteristics of CRM-induced occupational asthma. We encountered 16 cases of CRM-induced occupational asthma among farmers cultivating citrus fruits. Asthmatic attacks corresponded closely with their work on citrus farms. The mean duration of the latent period was 12.9 (range 7 to 20) years. During their first visit to our clinic, nine patients with FEV1 lower than 70% of predictive value showed reversible airway obstruction after inhalation of bronchodilator, and seven with FEV1 greater than 70% of predictive value showed airway hyperresponsiveness to methacholine. Fifteen of the 16 also complained of recurrent nasal symptoms, which had developed at an earlier time than the asthmatic symptoms. They showed strong positive reactions to CRM extract on skin prick test (A/H ratio > or = 1.0) and had high serum specific IgE antibody against CRM which was detected by ELISA. Skin prick test with common inhalant allergens revealed that 10 had an isolated positive response to CRM with negative results to common inhalant allergens in their environment. The ELISA inhibition tests with CRM demonstrated significant inhibitions by CRM in a dose-dependent manner, while minimal inhibitions were noted by D. pteronyssinus and mugwort allergens. These findings suggest that CRM could induce IgE-mediated bronchoconstriction in exposed workers on citrus farm.

  17. Overexpression of a Gene Encoding a Catabolite Repression Element in Alternaria citri Causes Severe Symptoms of Black Rot in Citrus Fruit.

    Science.gov (United States)

    Katoh, H; Ohtani, K; Yamamoto, H; Akimitsu, K

    2007-05-01

    ABSTRACT A gene (AcCreA) encoding a catabolite repression element (CreA) with (two zinc fingers of the Cys(2)His(2) type was isolated from the postharvest fungal pathogen Alternaria citri. The AcCreA overexpression mutant AcOEC2 of A. citri showed normal growth on pectin medium and on segments of peel or the juice sac area from citrus fruit. Production of endopolygalacturonase, an essential virulence factor of this pathogen, was similar in AcOEC2 and the wild type in pectin-containing media. However, addition of glucose to the medium showed that carbon catabolite repression of endopolygalacturonase gene (Acpg1) expression, as well as endopolygalacturonase production, was lost in AcOEC2. The wild-type strain of A. citri causes rot mainly in the central axis of citrus fruit without development of rotting in the juice sac area; however, AcOEC2 caused severe black rot symptoms in both the central axis and juice sac areas. These results indicate that AcCreA-mediated catabolite repression controls the virulence or infection of this pathogen, and that the wild-type A. citri does not cause symptoms in the juice sac area due to carbon catabolite repression by sugars in the juice of the juice sac area.

  18. Formation of Stylet Sheaths in aere (in air from eight species of phytophagous hemipterans from six families (Suborders: Auchenorrhyncha and Sternorrhyncha.

    Directory of Open Access Journals (Sweden)

    J Kent Morgan

    Full Text Available Stylet sheath formation is a common feature among phytophagous hemipterans. These sheaths are considered essential to promote a successful feeding event. Stylet sheath compositions are largely unknown and their mode of solidification remains to be elucidated. This report demonstrates the formation and solidification of in āere (in air produced stylet sheaths by six hemipteran families: Diaphorina citri (Psyllidae, Asian citrus psyllid, Aphis nerii (Aphididae, oleander/milkweed aphid, Toxoptera citricida (Aphididae, brown citrus aphid, Aphis gossypii (Aphididae, cotton melon aphid, Bemisia tabaci biotype B (Aleyrodidae, whitefly, Homalodisca vitripennis (Cicadellidae, glassy-winged sharpshooter, Ferrisia virgata (Pseudococcidae, striped mealybug, and Protopulvinaria pyriformis (Coccidae, pyriform scale. Examination of in āere produced stylet sheaths by confocal and scanning electron microscopy shows a common morphology of an initial flange laid down on the surface of the membrane followed by continuous hollow core structures with sequentially stacked hardened bulbous droplets. Single and multi-branched sheaths were common, whereas mealybug and scale insects typically produced multi-branched sheaths. Micrographs of the in āere formed flanges indicate flange sealing upon stylet bundle extraction in D. citri and the aphids, while the B. tabaci whitefly and H. vitripennis glassy-winged sharpshooter flanges remain unsealed. Structural similarity of in āere sheaths are apparent in stylet sheaths formed in planta, in artificial diets, or in water. The use of 'Solvy', a dissolvable membrane, for intact stylet sheath isolation is reported. These observations illustrate for the first time this mode of stylet sheath synthesis adding to the understanding of stylet sheath formation in phytophagous hemipterans and providing tools for future use in structural and compositional analysis.

  19. Differential expression of pathogenicity- and virulence-related genes of Xanthomonas axonopodis pv. citri under copper stress

    Directory of Open Access Journals (Sweden)

    Ana Carolina Basílio Palmieri

    2010-01-01

    Full Text Available In this study, we used real-time quantitative PCR (RT-qPCR to evaluate the expression of 32 genes of Xanthomonas axonopodis pv. citri related to pathogenicity and virulence that are also involved in copper detoxification. Nearly all of the genes were up-regulated, including copA and copB. Two genes homologous to members of the type II secretion system (xcsH and xcsC and two involved in the degradation of plant cell wall components (pglA and pel were the most expressed in response to an elevated copper concentration. The type II secretion system (xcs operon and a few homologues of proteins putatively secreted by this system showed enhanced expression when the bacteria were exposed to a high concentration of copper sulfate. The enhanced expression of the genes of secretion II system during copper stress suggests that this pathway may have an important role in the adaptative response of X. axonopodis pv. citri to toxic compounds. These findings highlight the potential role of these genes in attenuating the toxicity of certain metals and could represent an important means of bacterial resistance against chemicals used to control diseases.

  20. Functional analysis of a chitinase gene during the larval-nymph transition in Panonychus citri by RNA interference.

    Science.gov (United States)

    Xia, Wen-Kai; Shen, Xiao-Min; Ding, Tian-Bo; Niu, Jin-Zhi; Zhong, Rui; Liao, Chong-Yu; Feng, Ying-Cai; Dou, Wei; Wang, Jin-Jun

    2016-09-01

    Chitinases are hydrolytic enzymes that are required for chitin degradation and reconstruction in arthropods. In this study, we report a cDNA sequence encoding a putative chitinase (PcCht1) from the citrus red mite, Panonychus citri. The PcCht1 (564 aa) possessed a signal peptide, a conserver domain, and a chitin-binding domain. Structural and phylogenetic analyses found that PcCht1 had high sequence similarity to chitinases in Tetranychus urticae. Real-time quantitative PCR analyses showed that the transcript levels of PcCht1 peaked periodically in larval and nymph stages. Moreover, significant increase of PcCht1 transcript level in the larvae was observed upon the exposure of diflubenzuron. In contrast, exposures of the larvae to diflubenzuron resulted in the decreased chitin content. Furthermore, through a feeding-based RNA interference approach, we were able to reduce the PcCht1 transcript level by 59.7 % in the larvae, and consequently the treated larvae showed a very low molting rate compared with the control. Our results expanded the understanding of the important role of PcCht1 in the growth and development of P. citri.

  1. Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri

    Energy Technology Data Exchange (ETDEWEB)

    Santacruz, C. P.; Balan, A.; Ferreira, L. C. S. [Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP (Brazil); Barbosa, J. A. R. G., E-mail: joao@lnls.br [Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil); Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP (Brazil)

    2006-03-01

    The molybdate-binding protein (ModA) from X. axonopodis pv. citri was crystallized with sodium molybdate in the presence of PEG or sulfate. The crystal diffracted to a maximum resolution of 1.7 Å and belongs to the orthorhombic space group C222{sub 1,} with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. Xanthomonas axonopodis pv. citri ModA protein is the ABC periplasmic binding component responsible for the capture of molybdate. The protein was crystallized with sodium molybdate using the hanging-drop vapour-diffusion method in the presence of PEG or sulfate. X-ray diffraction data were collected to a maximum resolution of 1.7 Å using synchrotron radiation. The crystal belongs to the orthorhombic space group C222{sub 1}, with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress.

  2. Crystallographic structure and substrate-binding interactions of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri.

    Science.gov (United States)

    Balan, Andrea; Santacruz-Pérez, Carolina; Moutran, Alexandre; Ferreira, Luís Carlos Souza; Neshich, Goran; Gonçalves Barbosa, João Alexandre Ribeiro

    2008-02-01

    In Xanthomonas axonopodis pv. citri (Xac or X. citri), the modA gene codes for a periplasmic protein (ModA) that is capable of binding molybdate and tungstate as part of the ABC-type transporter required for the uptake of micronutrients. In this study, we report the crystallographic structure of the Xac ModA protein with bound molybdate. The Xac ModA structure is similar to orthologs with known three-dimensional structures and consists of two nearly symmetrical domains separated by a hinge region where the oxyanion-binding site lies. Phylogenetic analysis of different ModA orthologs based on sequence alignments revealed three groups of molybdate-binding proteins: bacterial phytopathogens, enterobacteria and soil bacteria. Even though the ModA orthologs are segregated into different groups, the ligand-binding hydrogen bonds are mostly conserved, except for Archaeglobus fulgidus ModA. A detailed discussion of hydrophobic interactions in the active site is presented and two new residues, Ala38 and Ser151, are shown to be part of the ligand-binding pocket.

  3. Antitussive, expectorant and anti-inflammatory activities of different extracts from Exocarpium Citri grandis.

    Science.gov (United States)

    Jiang, Kun; Song, Qian; Wang, Lei; Xie, Tianzhu; Wu, Xi; Wang, Ping; Yin, Guo; Ye, Wencai; Wang, Tiejie

    2014-10-28

    Exocarpium Citri grandis (C. grandis, Huajuhong in Chinese), the epicarp of C. grandis 'Tomentosa', is used as an antitussive, expectorant and anti-inflammatory drug for hundreds of years in China. The study was aimed at evaluating the antitussive, expectorant and anti-inflammatory effects of different extracts of C. grandis, providing experimental evidence for its traditional use, and laying a foundation for its further researches. Crude drugs of C. grandis were extracted with four kinds of solvents (water, 50% ethanol, 70% ethanol and 90% ethanol) in reflux conditions, respectively. Solutions were concentrated in reduced pressure and lyophilized in vacuum to yield the aqueous extract, 50% ethanolic extract, 70% ethanolic extract, and 90% ethanolic extract of C. grandis. Antitussive evaluations were carried out with ammonia liquor induced mice cough; expectorant effects were tested with phenol red secretion experiments in mice; anti-inflammatory effects were assessed by murine model of xylene induced ear edema in mice. Only aqueous and 70% ethanolic extracts of C. grandis displayed significant antitussive, expectorant and anti-inflammatory activities. Aqueous extract of C. grandis significantly decreased cough frequency caused by ammonia liquor, increased phenol red secretion and inhibited the development of ear edema in anti-inflammatory assay at the dose of 1005 mg/kg (Pstrong effect of decreasing cough frequency, prolonging cough period, increasing phenol red secretion and decreasing the extent of ear edema at the dose of 493 mg/kg (P<0.001). The low, middle, and high dose (247, 493, and 986 mg/kg) of 70% ethanolic extract of C. grandis showed significant antitussive, expectorant and anti-inflammatory effects in good dose dependant manner. The results supported the folk use of C. grandis (decoction of C. grandis) with scientific evidence, and indicated that the 70% ethanolic extract of C. grandis might have better effects of antitussive, expectorant and anti

  4. Localized autoinoculation and dissemination of Isaria fumosorosea for control of the Asian citrus psyllid in South Texas

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri Kuwayama, vectors the causal organism of citrus greening disease. Integrated strategies are needed to control D. citri in south Texas. Control approaches involving entomopathogenic fungi may be useful on ornamental and abandoned citrus and other rutaceous...

  5. Murraya paniculata (orange jasmine), a host and possible inoculum reservoir for Candidatus Liberibacter asiaticus, causal agent of Huanglongbing

    Science.gov (United States)

    Huanglongbing (HLB), considered the most serious vectored-pathogen of citrus, is transmitted in nature by the Asian citrus psyllid Diaphorina citri and the African citrus psyllid Trioza erytreae. In 1999, D. citri was discovered in southern Florida and has become established in FL and TX. Huanglon...

  6. The complete mitochondrial genome of the citrus red mite Panonychus citri (Acari: Tetranychidae): high genome rearrangement and extremely truncated tRNAs.

    Science.gov (United States)

    Yuan, Ming-Long; Wei, Dan-Dan; Wang, Bao-Jun; Dou, Wei; Wang, Jin-Jun

    2010-10-23

    The family Tetranychidae (Chelicerata: Acari) includes ~1200 species, many of which are of agronomic importance. To date, mitochondrial genomes of only two Tetranychidae species have been sequenced, and it has been found that these two mitochondrial genomes are characterized by many unusual features in genome organization and structure such as gene order and nucleotide frequency. The scarcity of available sequence data has greatly impeded evolutionary studies in Acari (mites and ticks). Information on Tetranychidae mitochondrial genomes is quite important for phylogenetic evaluation and population genetics, as well as the molecular evolution of functional genes such as acaricide-resistance genes. In this study, we sequenced the complete mitochondrial genome of Panonychus citri (Family Tetranychidae), a worldwide citrus pest, and provide a comparison to other Acari. The mitochondrial genome of P. citri is a typical circular molecule of 13,077 bp, and contains the complete set of 37 genes that are usually found in metazoans. This is the smallest mitochondrial genome within all sequenced Acari and other Chelicerata, primarily due to the significant size reduction of protein coding genes (PCGs), a large rRNA gene, and the A + T-rich region. The mitochondrial gene order for P. citri is the same as those for P. ulmi and Tetranychus urticae, but distinctly different from other Acari by a series of gene translocations and/or inversions. The majority of the P. citri mitochondrial genome has a high A + T content (85.28%), which is also reflected by AT-rich codons being used more frequently, but exhibits a positive GC-skew (0.03). The Acari mitochondrial nad1 exhibits a faster amino acid substitution rate than other genes, and the variation of nucleotide substitution patterns of PCGs is significantly correlated with the G + C content. Most tRNA genes of P. citri are extremely truncated and atypical (44-65, 54.1 ± 4.1 bp), lacking either the T- or D-arm, as found in P. ulmi

  7. The complete mitochondrial genome of the citrus red mite Panonychus citri (Acari: Tetranychidae: high genome rearrangement and extremely truncated tRNAs

    Directory of Open Access Journals (Sweden)

    Dou Wei

    2010-10-01

    Full Text Available Abstract Background The family Tetranychidae (Chelicerata: Acari includes ~1200 species, many of which are of agronomic importance. To date, mitochondrial genomes of only two Tetranychidae species have been sequenced, and it has been found that these two mitochondrial genomes are characterized by many unusual features in genome organization and structure such as gene order and nucleotide frequency. The scarcity of available sequence data has greatly impeded evolutionary studies in Acari (mites and ticks. Information on Tetranychidae mitochondrial genomes is quite important for phylogenetic evaluation and population genetics, as well as the molecular evolution of functional genes such as acaricide-resistance genes. In this study, we sequenced the complete mitochondrial genome of Panonychus citri (Family Tetranychidae, a worldwide citrus pest, and provide a comparison to other Acari. Results The mitochondrial genome of P. citri is a typical circular molecule of 13,077 bp, and contains the complete set of 37 genes that are usually found in metazoans. This is the smallest mitochondrial genome within all sequenced Acari and other Chelicerata, primarily due to the significant size reduction of protein coding genes (PCGs, a large rRNA gene, and the A + T-rich region. The mitochondrial gene order for P. citri is the same as those for P. ulmi and Tetranychus urticae, but distinctly different from other Acari by a series of gene translocations and/or inversions. The majority of the P. citri mitochondrial genome has a high A + T content (85.28%, which is also reflected by AT-rich codons being used more frequently, but exhibits a positive GC-skew (0.03. The Acari mitochondrial nad1 exhibits a faster amino acid substitution rate than other genes, and the variation of nucleotide substitution patterns of PCGs is significantly correlated with the G + C content. Most tRNA genes of P. citri are extremely truncated and atypical (44-65, 54.1 ± 4.1 bp, lacking

  8. Structural features of PhoX, one of the phosphate-binding proteins from Pho regulon of Xanthomonas citri

    Science.gov (United States)

    Pegos, Vanessa R.; Santos, Rodrigo M. L.; Medrano, Francisco J.

    2017-01-01

    In Escherichia coli, the ATP-Binding Cassette transporter for phosphate is encoded by the pstSCAB operon. PstS is the periplasmic component responsible for affinity and specificity of the system and has also been related to a regulatory role and chemotaxis during depletion of phosphate. Xanthomonas citri has two phosphate-binding proteins: PstS and PhoX, which are differentially expressed under phosphate limitation. In this work, we focused on PhoX characterization and comparison with PstS. The PhoX three-dimensional structure was solved in a closed conformation with a phosphate engulfed in the binding site pocket between two domains. Comparison between PhoX and PstS revealed that they originated from gene duplication, but despite their similarities they show significant differences in the region that interacts with the permeases. PMID:28542513

  9. Biocontrol ability and putative mode of action of yeasts against Geotrichum citri-aurantii in citrus fruit.

    Science.gov (United States)

    Ferraz, Luriany Pompeo; Cunha, Tatiane da; da Silva, Aline Caroline; Kupper, Katia Cristina

    2016-01-01

    Sour rot is a major postharvest disease of citrus fruit and is caused by the fungal pathogen Geotrichum citri-aurantii. A lack of chemicals certified for the control of this disease has led to the consideration of alternative methods and strategies, such as the use of yeasts as biocontrol agents. The purpose of the present study was to test the ability of yeasts isolated from leaves, flowers, fruit, and soil, and six Saccharomyces cerevisiae isolates to control citrus sour rot, to assess the mechanisms of action of the yeast isolates that were demonstrated to be effective for biocontrol, and to identify the most effective yeast isolates for the biocontrol of G. citri-aurantii. In in vivo assays, three yeast isolates (ACBL-23, ACBL-44, and ACBL-77) showed a potential for controlling sour rot in citrus fruits, both preventatively and curatively. Most of the eight yeast isolates that were assessed for a mechanism of action did not produce antifungal compounds in an amount sufficient to inhibit the growth of the pathogen. Additionally, nutrient competition among the yeast strains was not found to be a biocontrol strategy. Instead, killer activity and hydrolytic enzyme production were identified as the major mechanisms involved in the biocontrol activity of the yeasts. Isolates ACBL-23, ACBL-44, and ACBL-77, which controlled sour rot most effectively, were identified as Rhodotorula minuta, Candida azyma, and Aureobasidium pullulans, respectively. To our knowledge, this is the first report of the potential of C. azyma as a biological control agent against a postharvest pathogen and its ability to produce a killer toxin.

  10. Molecular Characterization of Vitellogenin and Its Receptor Genes from Citrus Red Mite, Panonychus citri (McGregor

    Directory of Open Access Journals (Sweden)

    Rui Zhong

    2015-03-01

    Full Text Available The production and uptake of yolk protein play an important role in the reproduction of all oviparous organisms. Vitellogenin (Vg is the precursor of vitellin (Vn, which is the major egg storage protein, and vitellogenin receptor (VgR is a necessary protein for the uptake of Vg into developing oocytes. In this paper, we characterize the full-length Vg and VgR, PcVg1 and PcVgR, respectively, of the citrus red mite Panonychus citri (McGregor. The PcVg1 cDNA is 5748 nucleotides (nt with a 5553-nt open reading frame (ORF coding for 1851 amino acids (aa, and the PcVgR is 6090 nt, containing an intact ORF of 5673 nt coding an expected protein of 1891 aa. The PcVg1 aa sequence shows a typical GLCG domain and several K/RXXR cleavage sites, and PcVgR comprises two ligand-binding domains, two epidermal growth factor (EGF-like regions containing YWTD motifs, a transmembrane domain, and a cytoplasmic domain. An analysis of the aa sequences and phylogenetics implied that both genes were genetically distinct from those of ticks and insects. The transcriptional profiles determined by real-time quantitative PCR in different developmental stages showed that both genes present the same expressional tendencies in eggs, larvae, nymphs, and adults. This suggested that the biosynthesis and uptake of PcVg occurs coordinately. The strong reproductive capacity of P. citri has been hypothesized as an important factor in its resistance; consequently, understanding the molecular mechanisms regulating Vg and VgR are fundamental for mite control.

  11. Monitoramento da expressão gênica de Xanthomonas axonopodis pv. citri em diferentes condições ambientais

    OpenAIRE

    2003-01-01

    The bacterium Xanthomonas axonopodis pv. citri is a pathogen causing citric canker that compromises part of the orange production in the State of São Paulo, with negative consequences for the economy of the State and the Country. For the future control of citric canker, several investigators participated in the Xanthomonas Genome Project in order to determine the gene set of this bacterium and to contribute to the identification of the genes involved in its pathogenicity. In this respect, the...

  12. Diel periodicity of pheromone release by females of Planococcus citri and Planococcus ficus and the temporal flight activity of their conspecific males

    Science.gov (United States)

    Levi-Zada, Anat; Fefer, Daniela; David, Maayan; Eliyahu, Miriam; Franco, José Carlos; Protasov, Alex; Dunkelblum, Ezra; Mendel, Zvi

    2014-08-01

    The diel periodicity of sex pheromone release was monitored in two mealybug species, Planococcus citri and Planococcus ficus (Hemiptera; Pseudococcidae), using sequential SPME/GCMS analysis. A maximal release of 2 ng/h pheromone by 9-12-day-old P. citri females occurred 1-2 h before the beginning of photophase. The highest release of pheromone by P. ficus females was 1-2 ng/2 h of 10-20-day-old females, approximately 2 h after the beginning of photophase. Mating resulted in termination of the pheromone release in both mealybug species. The temporal flight activity of the males was monitored in rearing chambers using pheromone baited delta traps. Males of both P. citri and P. ficus displayed the same flight pattern and began flying at 06:00 hours when the light was turned on, reaching a peak during the first and second hour of the photophase. Our results suggest that other biparental mealybug species display also diel periodicities of maximal pheromone release and response. Direct evaluation of the diel periodicity of the pheromone release by the automatic sequential analysis is convenient and will be very helpful in optimizing the airborne collection and identification of other unknown mealybug pheromones and to study the calling behavior of females. Considering this behavior pattern may help to develop more effective pheromone-based management strategies against mealybugs.

  13. Exposure to Diflubenzuron Results in an Up-Regulation of a Chitin Synthase 1 Gene in Citrus Red Mite, Panonychus citri (Acari: Tetranychidae

    Directory of Open Access Journals (Sweden)

    Wen-Kai Xia

    2014-02-01

    Full Text Available Chitin synthase synthesizes chitin, which is critical for the arthropod exoskeleton. In this study, we cloned the cDNA sequences of a chitin synthase 1 gene, PcCHS1, in the citrus red mite, Panonychus citri (McGregor, which is one of the most economically important pests of citrus worldwide. The full-length cDNA of PcCHS1 contains an open reading frame of 4605 bp of nucleotides, which encodes a protein of 1535 amino acid residues with a predicted molecular mass of 175.0 kDa. A phylogenetic analysis showed that PcCHS1 was most closely related to CHS1 from Tetranychus urticae. During P. citri development, PcCHS1 was constantly expressed in all stages but highly expressed in the egg stage (114.8-fold higher than in the adult. When larvae were exposed to diflubenzuron (DFB for 6 h, the mite had a significantly high mortality rate, and the mRNA expression levels of PcCHS1 were significantly enhanced. These results indicate a promising use of DFB to control P. citri, by possibly acting as an inhibitor in chitin synthesis as indicated by the up-regulation of PcCHS1 after exposure to DFB.

  14. Construction of EGFP-labeling system for visualizing the infection process of Xanthomonas axonopodis pv. citri in planta.

    Science.gov (United States)

    Liu, Li-Ping; Deng, Zi-Niu; Qu, Jin-Wang; Yan, Jia-Wen; Catara, Vittoria; Li, Da-Zhi; Long, Gui-You; Li, Na

    2012-09-01

    Xanthomonas axonopodis pv. citri (Xac) is the causal agent of citrus bacterial canker, an economically important disease to world citrus industry. To monitor the infection process of Xac in different citrus plants, the enhanced green florescent protein (EGFP) visualizing system was constructed to visualize the propagation and localization in planta. First, the wild-type Xac was isolated from the diseased leaves of susceptible 'Bingtang' sweet orange, and then the isolated Xac was labeled with EGFP by triparental mating. After PCR identification, the growth kinetics and pathogenicity of the transformants were analyzed in comparison with the wild-type Xac. The EGFP-labeled bacteria were inoculated by spraying on the surface and infiltration in the mesophyll of 'Bingtang' sweet orange leaves. The bacterial cell multiplication and diffusion processes were observed directly under confocal laser scanning microscope at different intervals after inoculation. The results indicated that the EGFP-labeled Xac releasing clear green fluorescence light under fluorescent microscope showed the infection process and had the same pathogenicity as the wild type to citrus. Consequently, the labeled Xac demonstrated the ability as an efficient tool to monitor the pathogen infection.

  15. The Effect of Ultraviolet-A Radiation Exposure on the Reproductive Ability, Longevity, and Development of the Dialeurodes citri (Homoptera: Aleyrodidae) F1 Generation.

    Science.gov (United States)

    Tariq, Kaleem; Noor, Mah; Saeed, Shafqat; Zhang, Hongyu

    2015-12-01

    Ultraviolet (UV) light has been used worldwide to monitor and trap insect pests. Whitefly adults show conspicuous positive phototactic behavior toward UV light stimuli; however, knowledge of the effect of UV light exposure on various life-history parameters of Dialeurodes citri remains limited. The present research aimed to investigate the effect of ultraviolet radiation (UV-A; long-wave) exposure on the reproduction and longevity of D. citri adults as well as the development of immature (eggs, larvae, and pupae) flies in the F1 generation. Paired D. citri adults were exposed to UV-A radiation for different periods (0, 1, 4, and 7 h/d) until the end of their life. The results of the experiment revealed that fecundity and oviposition rates increased when adults were irradiated for 1 and 4 h/d, but interestingly, both were significantly decreased compared with those of the controls after the longest exposure time (7 h/d). The longevity of adults of both sexes and the cumulative survival of F1 immatures were decreased with increased exposure time. Exposure to UV-A radiation prolonged the developmental time of immature stages, and a positive correlation was observed with exposure time. Exposure to UV light significantly inhibited egg hatching, larval development, pupation, and adult emergence. To the best of our knowledge, this is the first study describing the effect of UV radiation on a homopteran insect pest. This research may provide a foundation for the scientific community to use UV light in the field as an integrated pest management strategy to control this devastating agricultural pest.

  16. In silico identification of potential chaperone genes that belong to type III and type IV secretion systems in Xanthomonas axonopodis pv citri

    Directory of Open Access Journals (Sweden)

    Letícia Khater

    2005-01-01

    Full Text Available The secretion of bacterial virulence factors and flagellar components requires the assistance of specific type III and flagellar chaperones. Standard computational annotation of the genome of Xanthomonas axonopodis pv citri, a plant pathogen that causes citrus canker, initially did not identify any genes belonging to these chaperone categories since the primary sequence homology between them was very low. However, in a search for hypothetical proteins with characteristics similar to these chaperones, we have now identified 30 chromosomal and 10 plasmidial potential genes encoding chaperones belonging to types III/IV, and flagellar secretion systems in this organism. The significance of these findings is discussed.

  17. Biologia e tabela de vida de fertilidade de Planococcus citri em diferentes estruturas vegetativas de cultivares de videira Biology and fertility life table of Planococcus citri in different vegetative structures of grape cultivars

    Directory of Open Access Journals (Sweden)

    Wilson José Morandi Filho

    2008-08-01

    Full Text Available O objetivo deste trabalho foi estudar a biologia e a tabela de vida de fertilidade de Planococcus citri, em folhas de videira das cultivares Cabernet Sauvignon, Itália e Isabel, em bagas de uva 'Itália' e em raízes da 'Isabel' e dos porta-enxertos 101-14 e IAC-572. Os seguintes parâmetros foram avaliados: duração e viabilidade dos estágios de ovo e ninfa; e fecundidade e longevidade dos adultos. Em raízes do porta-enxerto IAC-572, a cochonilha não completou o ciclo biológico. Em bagas de uva 'Itália', o inseto completou a fase de ninfa, porém os adultos foram inférteis. Em folhas das diferentes cultivares, a duração média do período de ovo a adulto dos machos foi de 24,63 dias, com viabilidade de 32%, enquanto as fêmeas duraram 32 dias com viabilidade de 56%. Em raízes, a duração do ciclo biológico de fêmeas e machos foi de 32,45 e 29,50 dias, respectivamente. Em folhas, a fecundidade foi de 67,27, 66,09 e 53,33 ovos por fêmea, nas cultivares Cabernet Sauvignon, Isabel e Itália, respectivamente. Nas raízes foram obtidos 30,4 e 70,0 ovos por fêmea, no porta-enxerto 101-14 e na cultivar Isabel, respectivamente.The objective of this work was to study the biology and fertility life table of Planococcus citri in leaves of the grape cultivars Cabernet Sauvignon, Itália and Isabel, in berries of 'Itália', in roots of 'Isabel' and 101-14 and IAC-572 rootstocks. The following parameters were evaluated: duration and viability of the egg and nymph stages; and fertility and longevity of the adults. On berries of 'Itália', the insect completed the nymph phase, however the adults were infertile. On roots of the rootstock IAC-572, citrus mealybug was unable to develop. On leaves of the grape cultivars, the period of egg to adult males was 24.63 days, with viability of 32%, while females lasted 32 days with viability of 56%. On roots, the duration of the cycle of females and males was 32.45 and 29.50 days, respectively. For females

  18. Biologia do psilídeo-de-concha Glycaspis brimblecombei Moore (Hemiptera, Psyllidae em Eucalyptus spp. Red gum lerp psyllid Glycaspis brimblecombei Moore (Hemiptera, Psylidae biology in Eucalyptus spp.

    Directory of Open Access Journals (Sweden)

    Daniela Cristina Firmino-Winckler

    2009-01-01

    Full Text Available Biologia do psilídeo-de-concha Glycaspis brimblecombei Moore (Hemiptera, Psyllidae em Eucalyptus spp.. Glycaspis brimblecombei Moore também conhecido por psilídeo-de-concha, se caracteriza por ser uma espécie específica ao gênero Eucalyptus L'Her. Este trabalho teve por objetivo determinar o ciclo biológico de G. brimblecombei em Eucalyptus spp. O trabalho foi conduzido em câmara climatizada (BOD, sob a temperatura de 26 °C e fotofase de 12 horas. As espécies de Eucalyptus utilizadas para o experimento foram: Eucalyptus camaldulensis, E. tereticornis, E. urophylla, E. grandis, Corymbia citriodora e um híbrido de E. grandis x E. urophylla ('urograndis'. Inicialmente foram utilizados 100 repetições (ninfas tratamento (espécies de Eucalyptus. As avaliações foram diárias. Os parâmetros biológicos avaliados foram a duração e viabilidade do estágio ninfal, longevidade dos adultos, número de posturas/fêmea, duração do período embrionário, número e viabilidade ovos, longevidade dos adultos e duração do ciclo total. Os resultados obtidos demonstraram que salvo C. citriodora que apresentou ser letal ao desenvolvimento ninfal de G. brimblecombei as demais espécies de Eucalyptus testadas oferecem condições ao desenvolvimento biológico deste psilídeo, sendo que neste trabalho E. camaldulensis mostrou-se a mais adequada.Glycaspis brimblecombei Moore, also known as red gum lerp psyllid, is characterized to be specific to the genus Eucalyptus. This work aimed to evaluate G. brimblecombei biological cycle in Eucalyptus spp. The work was accomplished in acclimatized chamber (BOD, with temperature of 26 °C and photophase of 12 hours. Eucalyptus species used in this study were: Eucalyptus camaldulensis, E. tereticornis, E. urophylla, E. grandis, Corymbia citriodora and E grandis x E. urophylla hybrid ('urograndis'. Initially 100 replications (nymphs per treatment (Eucalyptus species were prepared. The evaluations were daily and

  19. Asian Citrus Psyllid Expression Profiles Suggest Candidatus Liberibacter Asiaticus-Mediated Alteration of Adult Nutrition and Metabolism, and of Nymphal Development and Immunity.

    Science.gov (United States)

    Vyas, Meenal; Fisher, Tonja W; He, Ruifeng; Nelson, William; Yin, Guohua; Cicero, Joseph M; Willer, Mark; Kim, Ryan; Kramer, Robin; May, Greg A; Crow, John A; Soderlund, Carol A; Gang, David R; Brown, Judith K

    2015-01-01

    The Asian citrus psyllid (ACP) Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the insect vector of the fastidious bacterium Candidatus Liberibacter asiaticus (CLas), the causal agent of citrus greening disease, or Huanglongbing (HLB). The widespread invasiveness of the psyllid vector and HLB in citrus trees worldwide has underscored the need for non-traditional approaches to manage the disease. One tenable solution is through the deployment of RNA interference technology to silence protein-protein interactions essential for ACP-mediated CLas invasion and transmission. To identify psyllid interactor-bacterial effector combinations associated with psyllid-CLas interactions, cDNA libraries were constructed from CLas-infected and CLas-free ACP adults and nymphs, and analyzed for differential expression. Library assemblies comprised 24,039,255 reads and yielded 45,976 consensus contigs. They were annotated (UniProt), classified using Gene Ontology, and subjected to in silico expression analyses using the Transcriptome Computational Workbench (TCW) (http://www.sohomoptera.org/ACPPoP/). Functional-biological pathway interpretations were carried out using the Kyoto Encyclopedia of Genes and Genomes databases. Differentially expressed contigs in adults and/or nymphs represented genes and/or metabolic/pathogenesis pathways involved in adhesion, biofilm formation, development-related, immunity, nutrition, stress, and virulence. Notably, contigs involved in gene silencing and transposon-related responses were documented in a psyllid for the first time. This is the first comparative transcriptomic analysis of ACP adults and nymphs infected and uninfected with CLas. The results provide key initial insights into host-parasite interactions involving CLas effectors that contribute to invasion-virulence, and to host nutritional exploitation and immune-related responses that appear to be essential for successful ACP-mediated circulative, propagative CLas transmission.

  20. Asian Citrus Psyllid Expression Profiles Suggest Candidatus Liberibacter Asiaticus-Mediated Alteration of Adult Nutrition and Metabolism, and of Nymphal Development and Immunity.

    Directory of Open Access Journals (Sweden)

    Meenal Vyas

    Full Text Available The Asian citrus psyllid (ACP Diaphorina citri Kuwayama (Hemiptera: Psyllidae is the insect vector of the fastidious bacterium Candidatus Liberibacter asiaticus (CLas, the causal agent of citrus greening disease, or Huanglongbing (HLB. The widespread invasiveness of the psyllid vector and HLB in citrus trees worldwide has underscored the need for non-traditional approaches to manage the disease. One tenable solution is through the deployment of RNA interference technology to silence protein-protein interactions essential for ACP-mediated CLas invasion and transmission. To identify psyllid interactor-bacterial effector combinations associated with psyllid-CLas interactions, cDNA libraries were constructed from CLas-infected and CLas-free ACP adults and nymphs, and analyzed for differential expression. Library assemblies comprised 24,039,255 reads and yielded 45,976 consensus contigs. They were annotated (UniProt, classified using Gene Ontology, and subjected to in silico expression analyses using the Transcriptome Computational Workbench (TCW (http://www.sohomoptera.org/ACPPoP/. Functional-biological pathway interpretations were carried out using the Kyoto Encyclopedia of Genes and Genomes databases. Differentially expressed contigs in adults and/or nymphs represented genes and/or metabolic/pathogenesis pathways involved in adhesion, biofilm formation, development-related, immunity, nutrition, stress, and virulence. Notably, contigs involved in gene silencing and transposon-related responses were documented in a psyllid for the first time. This is the first comparative transcriptomic analysis of ACP adults and nymphs infected and uninfected with CLas. The results provide key initial insights into host-parasite interactions involving CLas effectors that contribute to invasion-virulence, and to host nutritional exploitation and immune-related responses that appear to be essential for successful ACP-mediated circulative, propagative CLas

  1. Genome-wide mutagenesis of Xanthomonas axonopodis pv. citri reveals novel genetic determinants and regulation mechanisms of biofilm formation.

    Directory of Open Access Journals (Sweden)

    Jinyun Li

    Full Text Available Xanthomonas axonopodis pv. citri (Xac causes citrus canker disease, a major threat to citrus production worldwide. Accumulating evidence suggests that the formation of biofilms on citrus leaves plays an important role in the epiphytic survival of this pathogen prior to the development of canker disease. However, the process of Xac biofilm formation is poorly understood. Here, we report a genome-scale study of Xac biofilm formation in which we identified 92 genes, including 33 novel genes involved in biofilm formation and 7 previously characterized genes, colR, fhaB, fliC, galU, gumD, wxacO, and rbfC, known to be important for Xac biofilm formation. In addition, 52 other genes with defined or putative functions in biofilm formation were identified, even though they had not previously reported been to be associated with biofilm formation. The 92 genes were isolated from 292 biofilm-defective mutants following a screen of a transposon insertion library containing 22,000 Xac strain 306 mutants. Further analyses indicated that 16 of the novel genes are involved in the production of extracellular polysaccharide (EPS and/or lipopolysaccharide (LPS, 7 genes are involved in signaling and regulatory pathways, and 5 genes have unknown roles in biofilm formation. Furthermore, two novel genes, XAC0482, encoding a haloacid dehalogenase-like phosphatase, and XAC0494 (designated as rbfS, encoding a two-component sensor protein, were confirmed to be biofilm-related genes through complementation assays. Our data demonstrate that the formation of mature biofilm requires EPS, LPS, both flagellum-dependent and flagellum-independent cell motility, secreted proteins and extracellular DNA. Additionally, multiple signaling pathways are involved in Xac biofilm formation. This work is the first report on a genome-wide scale of the genetic processes of biofilm formation in plant pathogenic bacteria. The report provides significant new information about the genetic

  2. Effects of temperature on population growth parameters of Cryptolaemus montrouzieri (Coleoptera: Coccinellidae reared on Planococcus citri (Homoptera: Pseudococcidae

    Directory of Open Access Journals (Sweden)

    Negar Saeedi

    2015-09-01

    Full Text Available The effect of temperature on the development, survival, fecundity and population growth parameters of Cryptolaemus montrouzieri Mulsant (Col.: Coccinellidae reared on Planococcus citri Risso (Hom.: Pseudococcidae was determined at five constant temperatures (18, 22, 26, 30 and 32±1°C in the laboratory conditions. Results showed a temperature - dependent development of the coccinellid. The mean total of immature period was estimated 76.6±0.9, 33.9±0.1, 25.8±0.1, 22.6±0.1 and 26.2±1.4 days at 18, 22, 26, 30 and 32°C, respectively. The highest and lowest values of R0, rm and λ were obtained 369.9±50.2 and 2.5±0.8 (female/female/generation, 0.07±0.01 and 0.02±0.01 (female/female/day and 1.08±0.01 and 1.02±0.01 (day-1 at 26 and 32°C, respectively. The lowest values of generation time (T and doubling time (Dt were calculated 42.2±0.9 and 8.9±0.5 days at 32 and 26°C, respectively. The lower and higher developmental threshold of total of immature period were obtained 11.5 and 25.5°C, respectively. The thermal requirement for completion of total of immature period of this predator were estimated 400 DD. These results showed that 26°C and/or adjacent temperature is most suitable for mass rearing of this predator.

  3. Analysis of transcriptome differences between resistant and susceptible strains of the citrus red mite Panonychus citri (Acari: Tetranychidae.

    Directory of Open Access Journals (Sweden)

    Bin Liu

    Full Text Available BACKGROUND: The citrus red mite is a worldwide citrus pest and a common sensitizing allergen of asthma and rhinitis. It has developed strong resistance to many registered acaricides, However, the molecular mechanisms of resistance remain unknown. we therefore used next generation sequencing technology to investigate the global transcriptomes between resistant strains and susceptible strains. RESULTS: We obtained 34,159, 30,466 and 32,217 unigenes by assembling the SS reads, RS reads and SS&RS reads respectively. There are total 17,581 annotated unigenes from SS&RS reads by BLAST searching databases of nr, the Clusters of Orthologous Groups (COGs and Kyoto Encyclopedia of Genes and Genomes (KEGG with an E-value ≤ 1e-5, in which 7,075 unigenes were annotated in the COG database, 12, 712 unigenes were found in the KEGG database and 3,812 unigenes were assigned to Gene ontology (GO. Moreover, 2,701 unigenes were judged to be the differentially expressed genes (DEGs based on the uniquely mapped reads. There are 219 pathways in all annotated unigenes and 198 pathways in DEGs that mapped to the KEGG database. We identified 211 metabolism genes and target genes related to general insecticide resistance such as P450 and Cytochrome b, and further compared their differences between RS and SS. Meanwhile, we identified 105 and 194 genes related to growth and reproduction, respectively, based on the mode of action of Hexythiazox. After further analyses, we found variation in sequences but not in gene expression related to mite growth and reproduction between different strains. CONCLUSION: To our knowledge, this is the first comparative transcriptome study to discover candidate genes involved in phytophagous mite resistance. This study identified differential unigenes related to general pesticide resistance and organism growth and reproduction in P. citri. The assembled, annotated transcriptomes provide a valuable genomic resource for further understanding

  4. Suppression of greasy spot disease caused by Mycosphaerella citri Whiteside on grapefruit trees in an organic orchard using an aqueous organic mixture of composted cornmeal, humic acid, molasses, and fish oil vs vegetable oil

    Science.gov (United States)

    Greasy spot disease of citrus, caused by the fungus Mycosphaerella citri Whiteside, afflicts citrus trees in all citrus-growing areas of the United States, eastern Mexico, Central America, and the Caribbean islands, causing premature defoliation, blemished fruit, and reduced tree vigor, yield, and f...

  5. Assessment of Prey Preference by the Generalist Predator, Mallada basalis (Walker), When Offered Two Species of Spider Mites, Tetranychus kanzawai Kishida and Panonychus citri (McGregor) on Papaya

    Science.gov (United States)

    We investigated potential prey preference of the generalist predator Mallada basalis (Walker) when offered two mites, Tetranychus kanzawai Kishida and Panonychus citri (McGregor), both important pests on papaya. Laboratory choice tests revealed that none of the three larval instars of M. basalis sho...

  6. Characterization and functional analysis of a novel glutathione S-transferase gene potentially associated with the abamectin resistance in Panonychus citri (McGregor).

    Science.gov (United States)

    Liao, Chong-Yu; Xia, Wen-Kai; Feng, Ying-Cai; Li, Gang; Liu, Hai; Dou, Wei; Wang, Jin-Jun

    2016-09-01

    The citrus red mite, Panonychus citri (McGregor), a major citrus pest distributed worldwide, has been found to be resistant to various insecticides and acaricides used in China. However, the molecular mechanisms associated with the abamectin resistance in this species have not yet been reported. In this study, results showed over-expression of a novel glutathione S-transferases (GSTs) gene (PcGSTm5) in abamectin-resistant P. citri. Quantitative real-time PCR analysis showed that the transcripts of PcGSTm5 were also significantly up-regulated after exposure to abamectin and the maximum mRNA expression level at nymphal stage. The recombinant protein of PcGSTm5-pET-28a produced by Escherichia coli showed a pronounced activity toward the conjugates of 1-chloro-2,4 dinitrobenzene (CDNB) and glutathione (GSH). The kinetics of CDNB and GSH and its optimal pH and thermal stability were also determined. Reverse genetic study through a new method of leaf-mediated dsRNA feeding further support a link between the expression of PcGSTm5 and abamectin resistance. However, no direct evidence was found in metabolism or inhibition assays to confirm the hypothesis that PcGSTm5 can metabolize abamectin. Finally, it is here speculated that PcGSTm5 may play a role in abamectin detoxification through other pathway such as the antioxidant protection.

  7. New Protein-Protein Interactions Identified for the Regulatory and Structural Components and Substrates of the Type III Secretion System of the Phytopathogen Xanthomonas axonopodis Pathovar citri

    Science.gov (United States)

    Alegria, Marcos C.; Docena, Cassia; Khater, Leticia; Ramos, Carlos H. I.; da Silva, Ana C. R.; Farah, Chuck S.

    2004-01-01

    We have initiated a project to identify protein-protein interactions involved in the pathogenicity of the bacterial plant pathogen Xanthomonas axonopodis pv. citri. Using a yeast two-hybrid system based on Gal4 DNA-binding and activation domains, we have focused on identifying interactions involving subunits, regulators, and substrates of the type III secretion system coded by the hrp (for hypersensitive response and pathogenicity), hrc (for hrp conserved), and hpa (for hrp associated) genes. We have identified several previously uncharacterized interactions involving (i) HrpG, a two-component system response regulator responsible for the expression of X. axonopodis pv. citri hrp operons, and XAC0095, a previously uncharacterized protein encountered only in Xanthomonas spp.; (ii) HpaA, a protein secreted by the type III secretion system, HpaB, and the C-terminal domain of HrcV; (iii) HrpB1, HrpD6, and HrpW; and (iv) HrpB2 and HrcU. Homotropic interactions were also identified for the ATPase HrcN. These newly identified protein-protein interactions increase our understanding of the functional integration of phytopathogen-specific type III secretion system components and suggest new hypotheses regarding the molecular mechanisms underlying Xanthomonas pathogenicity. PMID:15342589

  8. Research of Exocarpium Citri Grandis Fingerprint Based on Self-organization Competitive Neural Network%基于自组织竞争型神经网络的化橘红指纹图谱研究

    Institute of Scientific and Technical Information of China (English)

    陈南迪; 卢丽萍; 汪金玉; 林励

    2012-01-01

    Objective To establish the self-organization competitive neural network method for identifying the species by the fingerprints of different kinds of Exocarpium Citri Grandis(Cirtus grandis Peel). Methods The fingerprints of different species of Exocarpium Citri Grandis were established by fourier transform infrared speetroscopy and high performance liquid chromatogram (HPLC). And then we used the self-organization competitive neural network model to distinguish the different species of Exocarpium Citri Grandis. Results The self-organization competitive neural network for distinguish Exocarpium Citri Grandis of different species was established, and the rate of accuracy for infrared and HPLC fingerprint of the powder and the extract was more than 91.67 % in average. Conclusion The self-organizing competitiveneural network can be used for distinguishing different species of Exocarpium Citri Grandis by their fingerprints.%目的 建立不同品种化橘红指纹图谱的自组织竞争型人工神经网络判别方法.方法 采用傅里叶红外光谱法及HPLC法建立不同品种化橘红的指纹图谱,采用自组织竞争型人工神经网络进行模式识别.结果 自组织竞争型神经网络模型对化橘红粉末、提取物红外及HPLC指纹图谱预测平均准确率达91.67%以上.结论 自组织竞争型人工神经网络可有效用于化橘红品种的识别.

  9. The monofunctional catalase KatE of Xanthomonas axonopodis pv. citri is required for full virulence in citrus plants.

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    Full Text Available BACKGROUND: Xanthomonas axonopodis pv. citri (Xac is an obligate aerobic phytopathogen constantly exposed to hydrogen peroxide produced by normal aerobic respiration and by the plant defense response during plant-pathogen interactions. Four putative catalase genes have been identified in silico in the Xac genome, designated as katE, catB, srpA (monofunctional catalases and katG (bifunctional catalase. METHODOLOGY/PRINCIPAL FINDINGS: Xac catalase activity was analyzed using native gel electrophoresis and semi-quantitative RT-PCR. We demonstrated that the catalase activity pattern was regulated in different growth stages displaying the highest levels during the stationary phase. KatE was the most active catalase in this phase of growth. At this stage cells were more resistant to hydrogen peroxide as was determined by the analysis of CFU after the exposition to different H(2O(2 concentrations. In addition, Xac exhibited an adaptive response to hydrogen peroxide, displaying higher levels of catalase activity and H(2O(2 resistance after treatment with sub-lethal concentrations of the oxidant. In the plant-like medium XVM2 the expression of KatE was strongly induced and in this medium Xac was more resistant to H(2O(2. A XackatE mutant strain was constructed by insertional mutagenesis. We observed that catalase induction in stationary phase was lost meanwhile the adaptive response to peroxide was maintained in this mutant. Finally, the XackatE strain was assayed in planta during host plant interaction rendering a less aggressive phenotype with a minor canker formation. CONCLUSIONS: Our results confirmed that in contrast to other Xanthomonas species, Xac catalase-specific activity is induced during the stationary phase of growth in parallel with the bacterial resistance to peroxide challenge. Moreover, Xac catalases expression pattern is modified in response to any stimuli associated with the plant or the microenvironment it provides. The catalase Kat

  10. Comparison of PCR,DIA and Pathogenicity Assay for Detection of Xanthomonas axonopodis pv.citri,the Causal Agent of Citrus Bacterial Canker Disease

    Institute of Scientific and Technical Information of China (English)

    WANG Zhong-kang; SUN Xian-yun; YIN You-ping; ZHOU Chang-yong; XIA Yu-xian

    2004-01-01

    Polymerase chain reaction (PCR) approach based on newly designed primers, JYF5/JYR5, was applied for specific detection of Xanthomonas axonopodis pv.citri(Xac). The efficiency and reliability of PCR method were compared with dot immunobinding assay (DIA) and classical pathogenicity test techniques for detecting suspensions of pure cells of Xac and soaking sap of citrus tissues. Detection sensitivity of PCR was about 4.5 cells or 1.56 pg target DNA per reaction which was higher than that of DIA (ca. 450 cells per dot).These three techniques (PCR assay, DIA and Pathogenecity test) could always detect Xac from symptomatic citrus samples. Different performances were obtained from citrus materials without symptoms, and the positive detection frequency was PCR, DIA and pathogenicity test.

  11. Xanthomonas axonopodis pv. citri YaeQ structure reveals new compact protein fold built around a variation of the PD-(D/E)XK nuclease motif

    Energy Technology Data Exchange (ETDEWEB)

    Guzzo, Cristiane Rodrigues; Farah, Chuck Shaker [Universidade de Sao Paulo (USP), SP (Brazil). Inst. de Quimica. Dept. de Bioquimica; Nagem, Ronaldo Alves Pinto [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Bioquimica e Imunologia; Barbosa, Joao Alexandre Ribeiro Goncalves [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil). Centro de Biologia Molecular Estrutural

    2008-11-15

    The YaeQ family of proteins derives from close to two hundred species of Gram-negative bacteria. Despite their widespread distribution, their molecular and physiological functions are unknown. We used X-ray crystallography and multi-wavelength anomalous dispersion to determine the high resolution structure of the YaeQ protein (XAC2398) from the phytopathogen Xanthomonas axonopodis pv. citri. The novel structure revealed that YaeQ pertains to the PD-(D/E)XK superfamily of metal-dependent endonucleases. The phylogenetic distribution of YaeQ proteins and a detailed comparison of the YaeQ structure with that of other PD-(D/E)XK family members point to specific testable hypotheses regarding YaeQ function. (author)

  12. Deciphering the regulon of a GntR family regulator via transcriptome and ChIP-exo analyses and its contribution to virulence in Xanthomonas citri.

    Science.gov (United States)

    Zhou, Xiaofeng; Yan, Qing; Wang, Nian

    2017-02-01

    Xanthomonas contains a large group of plant-associated species, many of which cause severe diseases on important crops worldwide. Six gluconate-operon repressor (GntR) family transcriptional regulators are predicted in Xanthomonas, one of which, belonging to the YtrA subfamily, plays a prominent role in bacterial virulence. However, the direct targets and comprehensive regulatory profile of YtrA remain unknown. Here, we performed microarray and high-resolution chromatin immunoprecipitation-exonuclease (ChIP-exo) experiments to identify YtrA direct targets and its DNA binding motif in X. citri ssp. citri (Xac), the causal agent of citrus canker. Integrative microarray and ChIP-exo data analysis revealed that YtrA directly regulates three operons by binding to a palindromic motif GGTG-N16 -CACC at the promoter region. A similar palindromic motif and YtrA homologues were also identified in many other bacteria, including Stenotrophomonas, Pseudoxanthomonas and Frateuria, indicating a widespread phenomenon. Deletion of ytrA in Xac abolishes bacterial virulence and induction of the hypersensitive response (HR). We found that YtrA regulates the expression of hrp/hrc genes encoding the bacterial type III secretion system (T3SS) and controls multiple biological processes, including motility and adhesion, oxidative stress, extracellular enzyme production and iron uptake. YtrA represses the expression of its direct targets in artificial medium or in planta. Importantly, over-expression of yro3, one of the YtrA directly regulated operons which contains trmL and XAC0231, induced weaker canker symptoms and down-regulation of hrp/hrc gene expression, suggesting a negative regulation in Xac virulence and T3SS. Our study has significantly advanced the mechanistic understanding of YtrA regulation and its contribution to bacterial virulence. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  13. 橘全爪螨有螨叶率防治指标的初步研究%Preliminary Study on the Controlling Index of the Leaf Incidence of Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    张志恒; 陈国庆; 陈亦根

    2000-01-01

    @@ 橘全爪螨(Panonychus citri Mcgregor)是为害柑橘的一种主要害虫,常因防治不及时而造成减产和品质下降.如何快速准确地实施虫情调查监测,适时指导药剂防治,是橘全爪螨综合治理的一个重要环节.

  14. All five host-range variants of Xanthomonas citri carry one pthA homolog with 17.5 repeats that determines pathogenicity on citrus, but none determine host-range variation.

    Science.gov (United States)

    Al-Saadi, Abdulwahid; Reddy, Joseph D; Duan, Yong P; Brunings, Asha M; Yuan, Qiaoping; Gabriel, Dean W

    2007-08-01

    Citrus canker disease is caused by five groups of Xanthomonas citri strains that are distinguished primarily by host range: three from Asia (A, A*, and A(w)) and two that form a phylogenetically distinct clade and originated in South America (B and C). Every X. citri strain carries multiple DNA fragments that hybridize with pthA, which is essential for the pathogenicity of wide-host-range X. citri group A strain 3213. DNA fragments that hybridized with pthA were cloned from a representative strain from all five groups. Each strain carried one and only one pthA homolog that functionally complemented a knockout mutation of pthA in 3213. Every complementing homolog was of identical size to pthA and carried 17.5 nearly identical, direct tandem repeats, including three new genes from narrow-host-range groups C (pthC), A(w) (pthAW), and A* (pthA*). Every noncomplementing paralog was of a different size; one of these was sequenced from group A* (pthA*-2) and was found to have an intact promoter and full-length reading frame but with 15.5 repeats. None of the complementing homologs nor any of the noncomplementing paralogs conferred avirulence to 3213 on grapefruit or suppressed avirulence of a group A* strain on grapefruit. A knockout mutation of pthC in a group C strain resulted in loss of pathogenicity on lime, but the strain was unaffected in ability to elicit an HR on grapefruit. This pthC- mutant was fully complemented by pthA, pthB, or pthC. Analysis of the predicted amino-acid sequences of all functional pthA homologs and nonfunctional paralogs indicated that the specific sequence of the 17th repeat may be essential for pathogenicity of X. citri on citrus.

  15. 湖南柑橘大实蝇对不同柑橘品种的危害特点研究初报%The Damage Characteristics of Tetradacus citri Chen on Different Citrus Varieties in Hunan Province

    Institute of Scientific and Technical Information of China (English)

    林文力; 杨水芝; 潘美山; 陈海玲; 黄泽培; 龙建国; 肖伏莲

    2011-01-01

    采用定点观察和普查的方式对湖南省吉首市、麻阳县柑橘大实蝇的危害特点进行了研究.结果表明:柑橘大实蝇对不同柑橘品种的危害时间由早到晚依次为:脐橙>温洲蜜柑>冰糖橙≥大红甜橙=广柑>椪柑;其危害最严重的品种是脐橙,虫果率达63%,其次是早熟温洲蜜柑和冰糖橙,达22%,再次是椪柑,达6%;同时还发现,柑橘不同品种果皮表面存在产卵假痕迹现象.这些结果可为柑橘大实蝇的防治提供理论参考依据.%The damage characteristics of Tetradacus citri Chen in Mayang County, Jishou City, Hunan Province were investigated by fixed-point observation method and general survey method. The results indicated that the harm time of Tetradacus citri Chen on different varieties of citrus ordered as navel orange>satsuma mandarin>Citrus sinensis ^Citrus sineasis Osbeck cv. Dahong= Citrussinensis (L.) Osbeck >Citrus reticulata Blanco cv. Ponkan; the damage level of Tetradacus citri Chen on different varieties of citrus ordered as navel orange>early mature satsuma mandarin and Citrus sinensis >dtrus reticulata Blanco cv. Ponkan, and the insect infestation ratio of navel orange, early mature satsuma mandarin, Citrus sinensis and Citrus reticulata Blanco cv. Ponkan was 63%, 22%, 22% and 6%, respectively. Meanwhile, the research also showed that there are fake oviposition traces of Tetradacus citri Chen on the Peels of different varieties of citrus. These results above can provide the theory basis for the control of Tetradacus citri Chen.

  16. 橘全爪螨种群空间分布与危害%Population Spacial Distribution and Harm of Panonychus citri

    Institute of Scientific and Technical Information of China (English)

    甘宗义; 朱红艳; 潭荣荣

    2012-01-01

    The spacial distribution pattern, population dynamics and harm of Panonychus citri McG on citrus tree were studied by test of theoretical distribution and analysis of spacial pattern indexes. The results showed that the population distribution of the mite suits with Negative-binomial distribution in citrus garden, and fits with Neyman distribution-A in lower population density too. The spacial distribution pattern presents loose colonies, in which distribution of the colonies is aggregated and the individuals in the colonies show random distribution. The aggregation intensity of population distribution is changing with changes in the mite population density. The aggregation in low density population is caused by environmental factors; and in highdeasity population it is concerned with their biological activities. When the population density a certain up to a certain harm grade, the spacial volume reaching and outstripping this harm level is not over 30% of the total space in citrus garden, accordingly, the area sprayed with insecticides for controlling the harmful mite should not be over 30% of all the area in the field.%通过理论分布的适合性检验、空间格局指标分析,对橘全爪螨(Panonychus citri McG)在温州蜜柑上的空间分布特征、动态与危害进行了系统研究.结果表明,橘全爪螨田间种群分布符合负二项分布,低密度下偶尔符合Neyman A分布.空间分布的基本成分为疏松的个体群,个体群内成员间的分布为随机分布,个体群间呈聚集分布,聚集强度随种群密度的变化而变化,种群低密度下的聚集主要由环境因素引起,高密度下的聚集与该螨自身生物学特性有关.当田间种群数量达到某一危害级别时,相应级别的危害空间容积不超过总体空间的30%,相应的田间药剂防治面积也不应超过总面积的30%.

  17. Occurrence and control of citrus melanose caused by Diaporthe citri%柑橘黑点病的发病规律和防治

    Institute of Scientific and Technical Information of China (English)

    姜丽英; 徐法三; 黄振东; 黄峰; 陈国庆; 李红叶

    2012-01-01

    近年来,柑橘黑点病(Diaporthe citri)在浙江、江西等柑橘产区普遍发生,逐渐成为柑橘最主要病害之一,严重影响鲜销果实的外观品质和销售价格.2008年以来,通过对该病的症状、发生规律和防治技术开展研究发现,果实整个生长期都能感病,但以果实膨大期最感病;黑点病的严重度与树龄、施肥水平和杀菌剂的使用次数关系密切;以座果后每隔20d使用保护性杀菌剂代森锰锌防治效果较好,代森锰锌中添加0.25%~0.50%矿物油(绿颖)能有效地提高其防效.文章同时结合国内外相关研究,对黑点病症状、病原、发生规律和防治技术作了较为详尽的综述.%Melanose, caused by Diaporthe citri, has become a serious disease on citrus fruits in citrus-growing region of Zhejiang, Jiangxi provinces etc. It deteriorates exterior quality and brings down sale price of citrus fruits for fresh marketing. We have been involved in the investigation of the symptoms, prevalence and control methods of this disease since 2008. It was demonstrated that citrus fruits are susceptible to this disease in whole developmental stage, and especially susceptible during the enlarging stage. The order of melanose severity was closely associated with citrus age, times of fertilization and fungicide application. The best control efficiency could be obtained by application of protective fungicides such as mancozeb (followed the instruction of manufactory) 20 days interval after fruit-setting. Superior efficiency would be even achieved when 0. 25% to 0. 5% mineral oil ( SK spray oil) was mixed with mancozeb. Inter- and intranational information regarding the symptoms, characteristics of pathogen, epidemic factors and disease control of melanose was elaborately integrated in this paper.

  18. Aspectos biológicos de Chrysoperla externa (Hagen, 1861(Neuroptera: Chrysopidae alimentada com Planococcus citri (Risso, 1813(Hemiptera: Pseudococcidae e Toxoptera citricida (Kirkaldy, 1907 (Hemiptera: Aphididae Bipogical aspects of Chrysoperla externa (Hagen, 1861 (Neuroptera: Chrysopidae fed with Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae and Toxoptera citricida (Kirkaldy, 1907 (Hemiptera: Aphididae (Hemiptera: Aphididae

    Directory of Open Access Journals (Sweden)

    Jean Patrick Bonani

    2009-02-01

    Full Text Available O ecossistema citrícola abriga muitas espécies de inimigos naturais, entre eles Chrysoperla externa (Hagen, 1861 que apresenta um grande potencial como agente de controle biológico de pragas. Neste trabalho, objetivou-se avapar o efeito das presas Planococcus citri (Risso, 1813 e Toxoptera citricida (Kirkaldy, 1907 sobre o desenvolvimento das fases de larva e pupa de C. externa. O ensaio foi conduzido em sala cpmatizada a uma temperatura de 25±1ºC, 70±10% UR e fotofase de 12 horas, sendo os tratamentos constituídos pelo fornecimento contínuo e alternado dessas duas presas em cada ínstar do predador, além de ovos de Anagasta kuehniella (Zeller, 1879, utilizados para fins de comparação. Foram avaliados o tempo de vida e a viabilidade das fases de larva, pupa e do período larva-adulto de C. externa. Constatou-se que T. citricida fornecido isoladamente ao longo do desenvolvimento larval ou em dois ínstares consecutivos do predador, não foi uma presa adequada, constatando-se 100% de mortalidade no período larva-adulto. Quando as larvas alimentaram-se de P. citri fornecida em dois ínstares do predador, a viabilidade no período larva-adulto foi semelhante à dieta constituída por apenas ovos de A. kuehniella, exceto quando se alimentaram de T. citricida no primeiro ínstar.The citrus-growing ecosystem houses a great deal of species of natural enemies, among them Chrysoperla externa (Hagen, 1861, which presents a great potential for use in the bipogical contrp of pests. The aim of this work was to evaluate the effects of the preys Planococcus citri (Risso, 1813 and Toxoptera citricida (Kirkaldy, 1907 on the development of larva and pupa of C. externa. The trial was carried out in an accpmatized room at 25±1ºC, 70±10% RH and 12-hour photophase, the treatments consisting of the continuous and alternate supply of those two preys in each instar of the predator, in apition to eggs of Anagasta kuehniella (Zeller, 1879 utilized for

  19. Factors affecting transmission rates of 'Candidatus Liberibacter asiaticus' by Asian citrus psyllid

    Science.gov (United States)

    The Asian citrus psyllid (ACP), Diaphorina citri, is an important pest because it transmits a bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) responsible for a serious disease of citrus known as Asiatic huanglongbing (citrus greening disease). USDA-ARS researchers recently established a program...

  20. Visualization of ‘Candidatus Liberibacter asiaticus’ cells in citrus seed coats with fluorescence in situ hybridization and transmission electron microscopy

    Science.gov (United States)

    ‘Candidatus Liberibacter asiaticus’ is bacterium implicated as the causal agent of the economically damaging disease of citrus called huanglongbing (HLB). The bacterium is spread by movement of infected citrus propagation material and by the Asian citrus psyllid, Diaphorina citri Kuwayama. Seed tr...

  1. Degradation products of citrus volatile organic compounds (VOCs) acting as phagostimulants that increase probing behavior of Asian citrus psyllid

    Science.gov (United States)

    Volatile phytochemicals play a role in orientation by phytophagous insects. We studied antennal and behavioral responses of the Asian citrus psyllid, Diaphorina citri Kuwayama, vector of the citrus greening disease pathogen. Little or no response to citrus leaf volatiles was detected by electroanten...

  2. Trailers transporting oranges to processing plants move Asian citrus psyllids

    Science.gov (United States)

    Huanglongbing (citrus greening) is one of the most serious of citrus diseases. Movement of the disease occurs as a result of natural vector-borne infection and by movement of plant material. We demonstrate here that Diaphorina citri Kuwayama (vector of citrus greening pathogens) can be transported i...

  3. Incidence of ‘Candidatus Liberibacter asiaticus’ in a Florida population of Asian citrus psyllid

    Science.gov (United States)

    A study was conducted to assess the incidence of a bacterium ‘Candidatus Liberibacter asiaticus’ in a Florida population of Asian citrus psyllid (ACP), Diaphorina citri. The bacterium is the presumed causal agent of Asiatic huanglongbing, a serious citrus disease also known as citrus greening or yel...

  4. Dicty_cDB: SFB147 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 5 DQ673411_1( DQ673411 |pid:none) Diaphorina citri putative ribosoma... 48 9e-05 EU934275_1( EU934275 |pid:none) TSA: Anopheles darli...ngi AD-251 40S... 48 9e-05 EF634885_1( EF634885 |pid:none) Pristionchus sp. 10 RS51

  5. Broad sprectrum potential of Isaria fumosorosea on insect pests of citrus

    Science.gov (United States)

    Use of entomopathogenic fungi, Isaria fumosorosea, Ifr, =Paecilomyces fumosoroseus, successfully increased insect pest mortality. Spraying the Ifr containing product, PFR97 TM, on citrus seedlings was used to screen efficacy for the management of Asian citrus psyllid, Diaphorina citri; glassy-winge...

  6. Correlation of electronic monitoring and stylet pathways elucidate the role of sclerenchymatous ring as a barrier to phloem feeding on citrus leaves by Asian citrus psyllid

    Science.gov (United States)

    Asian citrus psyllid (ACP, Diaphorina. citri) feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas) bacterium that causes the economically devastating citrus greening disease. Recent studies have shown a fibrous ring of thick-wal...

  7. PROCENTAJE DE PSILIDOS PORTADORES DE Canidatus Liberibacter asiaticus Y SU RELACION CON LA INCIDENCIA DE HLB EN CINCO HUERTAS COMERCIALES DE LIMON MEXICANO {Citrus aurantifolia (Christm) Swingle} EN EL ESTADO DE COLIMA MEXICO

    Science.gov (United States)

    Asian citrus psyllids (Diaphorina citri) were individually analyzed by qPCR to detect Candidatus Liberibacter asiaticus (CLas). The psyllids were collected in Mexican lime (Citrus aurantifolia) trees in five commercial orchards in Tecomán and Manzanillo, Colima with severe symptoms of classical mott...

  8. Candidatus Liberibacter asiaticus is present in orange jasmine and Asian citrus psyllid reared from jasmine at low titers

    Science.gov (United States)

    Orange jasmine, Murraya paniculata, is a common horticultural plant in Florida, and an alternate host of the Asian citrus psyllid, Diaphorina citri Kuwayama. Orange jasmine has also been reported to harbor the bacteria Candidatus Liberibacter asiaticus, the causal agent of huanglongbing disease. We ...

  9. Bionomics of Asian citrus psyllid (Hemiptera: Liviidae) associated with orange jasmine hedges in southest central Florida, with special reference to biological control by Tamarixia radiata

    Science.gov (United States)

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is an important pest in Florida because it vectors bacteria responsible for citrus huanglongbing disease. In addition to infesting citrus, orange jasmine (Murraya paniculata) is one of ACP’s favorite host plants and is widely grown as an orn...

  10. Competition between honeydew producers in an ant-hemipteran interaction may enhance biological control of an invasive pest.

    Science.gov (United States)

    Tena, A; Hoddle, C D; Hoddle, M S

    2013-12-01

    Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an invasive citrus pest in southern California, which secretes honeydew and has the potential to spread a lethal bacterial disease, huanglongbing, of citrus. In urban citrus, Argentine ant, Linepithema humile (Mayr) (Hymenoptera: Formicidae), also an invasive pest, tends honeydew-producing hemipterans. We used field data to determine whether the mutualistic relationship between L. humile and six established species of honeydew producers may hinder or favor the establishment of D. citri and its biological control with Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae) in citrus via competition or mutualism for ants, respectively. In the field, L. humile and D. citri are engaged in a mutualistic relationship. Ants harvest solid honeydew secreted by psyllid nymphs and tended more than 55% of observed D. citri colonies. Linepithema humile displayed a preference hierarchy when tending honeydew producers infesting citrus. It responded equally or less intensively to D. citri than to other honeydew-producing species. Consequently, the mutualism between L. humile and D. citri was affected by the presence of other honeydew-producing species, and the percentage of D. citri colonies tended by L. humile. The number of ants per D. citri colony also decreased as the number of other honeydew producers increased. Diaphorina citri density was also affected by the presence of other honeydew producers. Both colony size and the number of D. citri nymphs counted per tree decreased as the number of other honeydew producers increased. Our results indicate that competition between honeydew producers for the mutualist ant L. humile may hinder the establishment of D. citri by possibly facilitating increased biological control.

  11. Repellent Activity of Extracts of Wild Rice Species against Panonychus citri and Aphis citricola in Associated with Esterase Isoenzyme in Insests

    Institute of Scientific and Technical Information of China (English)

    WAN Shu-qing; LIU Xiang-fa; FENG Guo-zhong; PAN Da-jian

    2006-01-01

    Six species of wild rice with different ecophenotypes including Oryza grandiglumis (E6-1, E6-3 / 6-4), O. minuta (E13-9,E13-13), O. officinalis (E15-8, E15-13), O. punctata (E16-1, E16-3, E1 6-13), O. granulata (E7-4), and O. latifolia (101392, E9-1, E9-10)were extracted with methnol and the repellent activity of the extracts against the two insects Aphis citricola and Panonychus citri were studied. The extracts of O. officinalis E15-8 showed higher repellent rate to the two insects than those of the other species. The repellent rates of the extracts of E15-8 to P. citriand A. citricola were 83.26% and 87.86% at 5×104 μg/mL in 24 h and 87.95% and 82.43% in 48 h, respectively. The extracts of O. officinalis E15-8 had the effect of inhibition to the esterase of the two insects.

  12. The filamentous phage XacF1 causes loss of virulence in Xanthomonas axonopodis pv. citri, the causative agent of citrus canker disease

    Directory of Open Access Journals (Sweden)

    Abdelmonim Ali Ahmad

    2014-07-01

    Full Text Available In this study, filamentous phage XacF1, which can infect Xanthomonas axonopodis pv. citri (Xac strains, was isolated and characterized. Electron microscopy showed that XacF1 is a member of the family Inoviridae and is about 1000 nm long and 16 nm in width. The genome of XacF1 is 7325 nucleotides in size, containing 13 predicted open reading frames (ORFs, some of which showed significant homology to Ff-like phage proteins such as ORF1 (pII, ORF2 (pV, ORF6 (pIII, and ORF8 (pVI. XacF1 showed a relatively wide host range, infecting seven out of 11 strains tested in this study. Frequently, XacF1 was found to be integrated into the genome of Xac strains. This integration occurred at the host dif site (attB and was mediated by the host XerC/D recombination system. The attP sequence was identical to that of Xanthomonas phage Cf1c. Interestingly, infection by XacF1 phage caused several physiological changes to the bacterial host cells, including lower levels of extracellular polysaccharide production, reduced motility, slower growth rate, and a dramatic reduction in virulence. In particular, the reduction in virulence suggested possible utilization of XacF1 as a biological control agent against citrus canker disease.

  13. The weathervane model, a functional and structural organization of the two-component alkanesulfonate oxidoreductase SsuD from Xanthomonas citri

    Energy Technology Data Exchange (ETDEWEB)

    Pegos, V.R. [Universidade Estadual de Campinas (UNICAMP), SP (Brazil); Oliveira, P.S.L.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil)

    2012-07-01

    Full text: In Xanthomonas citri, the phytopathogen responsible for the canker citrus disease, we identified in the ssuABCDE operon, genes encoding the alkanesulfonate ABC transporter as well as the two enzymes responsible for oxido reduction of the respective substrates. SsuD and SsuE proteins represent a two-component system that can be assigned to the group of FMNH{sub 2} -dependent monooxygenases. How- ever, despite of the biochemical information about SsuD and SsuE orthologs from Escherichia coli, there is no structural information of how the two proteins work together. In this work, we used ultracentrifugation, SAXS data and molecular modeling to construct a structural/functional model, which consists of eight molecules organized in a weathervane shape. Through this model, SsuD ligand-binding site for NADPH{sub 2} and FMN substrates is clearly exposed, in a way that might allow the protein-protein interactions with SsuE. Moreover, based on molecular dynamics simulations of SsuD in apo state, docked with NADPH{sub 2}, FMN or both substrates, we characterized the residues of the pocket, the mechanism of substrate interaction and transfer of electrons from NADPH{sub 2} to FMN. This is the first report that links functional and biochemical data with structural analyses. (author)

  14. Evaluation of suitable reference genes for quantitative RT-PCR during development and abiotic stress in Panonychus citri (McGregor) (Acari: Tetranychidae).

    Science.gov (United States)

    Niu, Jin-Zhi; Dou, Wei; Ding, Tian-Bo; Yang, Li-Hong; Shen, Guang-Mao; Wang, Jin-Jun

    2012-05-01

    Quantitative real time reverse transcriptase polymerase chain reaction (RT-qPCR) is preferred for gene expression analysis in living organisms. Currently, it is a valuable tool for biological and ecological studies as it provides a relatively straightforward way to assess the relevance of transcriptional regulation under developmental and stress tolerance conditions. However, studies have shown that some commonly used reference genes varied among different experimental treatments, thus, systematic evaluation of reference genes is critical for gene expression profiling, which is often neglected in gene expression studies of arthropods. The aim of this study is to identify the suitable reference genes for RT-qPCR experiments involving various developmental stages and/or under abiotic stresses in citrus red mite Panonychus citri, a key pest in citrus orchards worldwide. GeNorm, NormFinder, and Bestkeeper software analysis indicates that elongation factor-1 alpha (ELF1A), RNA polymerase II largest subunit, alpha tublin, and glyceraldhyde-3-phosphate dehydrogenase (GAPDH) are the most stable reference genes in various developmental stages, meanwhile, ELF1A and GAPDH were the most stable reference genes under various abiotic stresses. Furthermore, this study will serve as a resource to screen reference genes for gene expression studies in any other spider mite species.

  15. Optimization of Extraction Technology for Inositol from Exocarpium Citri Grandis%正交试验法优选化橘红中肌醇提取工艺

    Institute of Scientific and Technical Information of China (English)

    陈地灵; 陈振兴; 林励; 吴泽青; 吴俊生

    2012-01-01

    [目的]优选化橘红中肌醇最佳提取工艺.[方法]以乙醇浓度、料液比和提取温度为考察因素,采用高效液相一蒸发光散射检测器(HPLC-ELSD)色谱法测定化橘红中肌醇含量,采用正交试验法优选化橘红中肌醇最优提取工艺.[结果]化橘红中肌醇最佳提取工艺为体积分数70%乙醇、料液比1∶20、提取温度100℃、每次时间45 min、提取2次.[结论]本研究优选得到的提取工艺稳定、提取率高,可用于化橘红中肌醇的提取.%Objective To optimize the extraction technology for the inositol from Exocarpium Citri Grandis. Methods With ethanol concentration, the ratio of material to solvent and extracting temperature as factors, the extraction technology for the inositol from Exocarpium Citri Crandis was optimized by orthogonal design and through the determination of inositol content by high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD). Results The optimal extraction technology was as follows; 70% ethanol, the ratio of material to solvent being 1:20, extraction at 100℃, for 45 min and extraction twice. Conclusion The extraction technology is steady, extraction percentage is higher and the technology is suitable for the extraction of inositol from Exocarpium Citri Crandis.

  16. Studies on Growth and Decline Law and Chemical Control of Panonychus citri in Lemon Orchard in Ruili%瑞丽柠檬园红蜘蛛消长规律及药剂防治研究

    Institute of Scientific and Technical Information of China (English)

    王自然; 段惠芬; 彭满秀; 郭俊; 赵俊; 潘黎; 高俊燕

    2014-01-01

    The growth and decline law and chemical control of Panonychus citri in lemon orchard are studied in Ruili city , Yunnan province .The results indicate that Panonychus citri occurs mainly from November to the next January , and its population density reaches a peak (2500~3000 heads per plant) in middle January;the occurrence quantity of this mite is more in the north and south of lemon crown;the occurrence and damage of Panonychus citri in lemon orchard are closely related with temperature , hu-midity, food, fertilization, chemical control and so on;among the tested 7 kinds of pesticides, 4%Spirodiclofen SC, 1.8%Abam-ectin EC and 73%Propargite EC have better control effects on Panonychus citri, and they are safe and harmless to the young shoots , flowers and young fruits of lemon .%在云南省瑞丽市进行了柠檬园红蜘蛛的消长规律及药剂防治研究。结果表明:柠檬红蜘蛛主要在11月至次年1月发生,其发生量在1月中旬达到高峰(2500~3000头/株);红蜘蛛在树冠北方、南方的发生量较多;柠檬园红蜘蛛的发生和为害与温度、湿度、食料、施肥、施药等因素关系密切;在7种供试药剂中,以4%螺螨酯悬浮剂、1.8%阿维菌素乳油和73%炔螨特乳油对红蜘蛛的防治效果较好,且对柠檬的嫩梢、花和幼果安全无害。

  17. 基于电子舌技术对不同年份的化橘红的识别%Discrimination of Exocarpium Citri Grandis in Different Years Based on Electronic Tongue Techniques

    Institute of Scientific and Technical Information of China (English)

    熊萧萧; 王鲁峰; 徐晓云; 潘思轶; 王可兴

    2012-01-01

    不同年份的化橘红功效和价格均存在差异,为了快速识别未知橘红样品的年份,采用电子舌对2001~2010年的10个样品和一个盲样进行了分析.对原始数据采用主成分分析和判别因子分析.结果表明:主成分分析可以将不同年份的样品进行有效的区分,判别因子分析可以更好地区分不同年份的样品,并能进行盲样识别.在模式识别中,判别因子分析法的识别结果均优于主成分分析法,前者2个判别因子的累计方差贡献量达到了99.929%.电子舌可以较好地区分不同年份的化橘红,改进后有望应用于化橘红的快速检测.%The effect and price vary in Exocarpium Citri Grandis of different years. Ten samples of Exocarpium Citri Grandis produced from 2001 to 2010 were analyzed with electronic tongue to discriminate their ages. The study shows that both Principal Component Analysis (PCA) and Discriminant Factor Analysis (DFA) can be used to discriminate samples of different years with the latter mathod being more accurate. In the pattern recognition, the result of DFA is also more accurate than PCA with the fact that its conmulative contribution of 2-dimentional principal component reached as high as 99.929%. In conclusion, electronic tongue can be utilized as an effective way to discriminate the age level of Exocarpium Citri Grandis. The results of this study suggest the possibility of application of the propsed method in the rapid detection and analysis of Exocarpium Citri Grandis in the process of production, sales, and other related fields.

  18. Efeito do meio de cultura, temperatura, fotoperíodo e fungicidas no crescimento micelial e no controle de Alternaria alternata f. sp. citri, causador da mancha marrom do tangor murcote

    OpenAIRE

    Colturato,Adimara Bentivoglio

    2006-01-01

    A mancha marrom de alternaria, causada pela Alternaria alternata f. sp. citri, afeta o tangor murcote causando lesões em folhas, ramos e frutos, acarretando em desfolha e manchas que depreciam o fruto comercialmente. Os objetivos deste trabalho foram verificar o efeito do meio de cultura no isolamento, da temperatura e do fotoperíodo no crescimento micelial do patógeno, avaliar a sensibilidade do patógeno à fungicidas in vitro e estabelecer o melhor produto e dose para o controle da doença no...

  19. Wolbachia infection density in populations of the Asian citrus psyllid (Hemiptera: Liviidae).

    Science.gov (United States)

    Hoffmann, M; Coy, M R; Kingdom Gibbard, H N; Pelz-Stelinski, K S

    2014-10-01

    The symbiotic relationships between bacteria of the genus Wolbachia (order Rickettsiales) and their arthropod hosts are diverse and can range from mutualism to parasitism. Whereas effects of Wolbachia on host biology are well investigated, little is known about diversity and abundance of Wolbachia in their natural hosts. The phloem-feeding Asian citrus psyllid, Diaphorina citri (Kuwayama) (Hemiptera: Liviidae), is naturally infected with Wolbachia (wDi). In the current study, we calculated the within-host density of Wolbachia in Florida D. citri populations using quantitative polymerase chain reaction for detection of the Wolbachia outer surface protein gene, wsp. Gene quantities were normalized to the D. citri wingless gene (Wg) to estimate Wolbachia abundance in individual D. citri. Using this method, significant geographic differences in Wolbachia densities were detected among Florida D. citri populations, with higher infection levels occurring in male versus female hosts.

  20. Development and validation of a Xanthomonas axonopodis pv. citri DNA microarray platform (XACarray generated from the shotgun libraries previously used in the sequencing of this bacterial genome

    Directory of Open Access Journals (Sweden)

    Zaini Paulo A

    2010-05-01

    Full Text Available Abstract Background From shotgun libraries used for the genomic sequencing of the phytopathogenic bacterium Xanthomonas axonopodis pv. citri (XAC, clones that were representative of the largest possible number of coding sequences (CDSs were selected to create a DNA microarray platform on glass slides (XACarray. The creation of the XACarray allowed for the establishment of a tool that is capable of providing data for the analysis of global genome expression in this organism. Findings The inserts from the selected clones were amplified by PCR with the universal oligonucleotide primers M13R and M13F. The obtained products were purified and fixed in duplicate on glass slides specific for use in DNA microarrays. The number of spots on the microarray totaled 6,144 and included 768 positive controls and 624 negative controls per slide. Validation of the platform was performed through hybridization of total DNA probes from XAC labeled with different fluorophores, Cy3 and Cy5. In this validation assay, 86% of all PCR products fixed on the glass slides were confirmed to present a hybridization signal greater than twice the standard deviation of the deviation of the global median signal-to-noise ration. Conclusions Our validation of the XACArray platform using DNA-DNA hybridization revealed that it can be used to evaluate the expression of 2,365 individual CDSs from all major functional categories, which corresponds to 52.7% of the annotated CDSs of the XAC genome. As a proof of concept, we used this platform in a previously work to verify the absence of genomic regions that could not be detected by sequencing in related strains of Xanthomonas.

  1. A plant natriuretic peptide-like molecule of the pathogen Xanthomonas axonopodis pv. citri causes rapid changes in the proteome of its citrus host

    Directory of Open Access Journals (Sweden)

    Ottado Jorgelina

    2010-03-01

    Full Text Available Abstract Background Plant natriuretic peptides (PNPs belong to a novel class of peptidic signaling molecules that share some structural similarity to the N-terminal domain of expansins and affect physiological processes such as water and ion homeostasis at nano-molar concentrations. The citrus pathogen Xanthomonas axonopodis pv. citri possesses a PNP-like peptide (XacPNP uniquely present in this bacteria. Previously we observed that the expression of XacPNP is induced upon infection and that lesions produced in leaves infected with a XacPNP deletion mutant were more necrotic and lead to earlier bacterial cell death, suggesting that the plant-like bacterial PNP enables the plant pathogen to modify host responses in order to create conditions favorable to its own survival. Results Here we measured chlorophyll fluorescence parameters and water potential of citrus leaves infiltrated with recombinant purified XacPNP and demonstrate that the peptide improves the physiological conditions of the tissue. Importantly, the proteomic analysis revealed that these responses are mirrored by rapid changes in the host proteome that include the up-regulation of Rubisco activase, ATP synthase CF1 α subunit, maturase K, and α- and β-tubulin. Conclusions We demonstrate that XacPNP induces changes in host photosynthesis at the level of protein expression and in photosynthetic efficiency in particular. Our findings suggest that the biotrophic pathogen can use the plant-like hormone to modulate the host cellular environment and in particular host metabolism and that such modulations weaken host defence.

  2. Effect of temperature on life history and population growth parameters of Planococcus citri (Homoptera, Pseudococcidae on coleus [Solenostemon scutellarioides (L. Codd.

    Directory of Open Access Journals (Sweden)

    Goldasteh Shila

    2009-01-01

    Full Text Available The development, life history, reproduction, and population growth parameters of Planococcus citri Risso on coleus [Solenostemon scutellarioides (L. Codd.] were studied at various temperatures ranging from 10 to 37ºC, 70±10% RH, and photoperiod length of 16: 8 h (L: D. Females and males successfully developed into adults at from 15 to 32ºC and 18 to 32ºC, respectively. All first instars died at 10, 12, and 37ºC. Lower temperatures (10, 12, and 15ºC caused higher egg mortality than did higher temperatures (32, 35, and 37ºC. At all temperatures (except 15ºC, the highest percentage of nymphal mortality was observed in the first instar. The sex ratio was female-biased between 15 and 30ºC, but there was a slightly higher number of males at 32ºC. The highest adult longevities of females and males were obtained at 18 and 25°C, respectively. The pre-oviposition, oviposition, and post-oviposition periods were significantly different at various temperatures. The highest fecundity was observed at 23ºC. The shortest and longest oviposition periods occurred at 32 and 18ºC, respectively. Maximum values of the intrinsic rate of natural increase (rm, net reproduction rate (R0, and finite rate of increase (λ and the shortest mean generation time (T and doubling time (DT were obtained at 25ºC. Our findings showed citrus mealybug performances to be highly affected by temperature.

  3. A plant natriuretic peptide-like molecule of the pathogen Xanthomonas axonopodis pv. citri causes rapid changes in the proteome of its citrus host

    KAUST Repository

    Garavaglia, Betiana S

    2010-03-21

    Background: Plant natriuretic peptides (PNPs) belong to a novel class of peptidic signaling molecules that share some structural similarity to the N-terminal domain of expansins and affect physiological processes such as water and ion homeostasis at nano-molar concentrations. The citrus pathogen Xanthomonas axonopodis pv. citri possesses a PNP-like peptide (XacPNP) uniquely present in this bacteria. Previously we observed that the expression of XacPNP is induced upon infection and that lesions produced in leaves infected with a XacPNP deletion mutant were more necrotic and lead to earlier bacterial cell death, suggesting that the plant-like bacterial PNP enables the plant pathogen to modify host responses in order to create conditions favorable to its own survival.Results: Here we measured chlorophyll fluorescence parameters and water potential of citrus leaves infiltrated with recombinant purified XacPNP and demonstrate that the peptide improves the physiological conditions of the tissue. Importantly, the proteomic analysis revealed that these responses are mirrored by rapid changes in the host proteome that include the up-regulation of Rubisco activase, ATP synthase CF1 ? subunit, maturase K, and ?- and ?-tubulin.Conclusions: We demonstrate that XacPNP induces changes in host photosynthesis at the level of protein expression and in photosynthetic efficiency in particular. Our findings suggest that the biotrophic pathogen can use the plant-like hormone to modulate the host cellular environment and in particular host metabolism and that such modulations weaken host defence. 2010 Garavaglia et al; licensee BioMed Central Ltd.

  4. Rapid screening and identification of compounds with DNA-binding activity from Folium Citri Reticulatae using on-line HPLC-DAD-MS(n) coupled with a post column fluorescence detection system.

    Science.gov (United States)

    Fu, Qingrong; Zhang, Cangman; Lin, Zongtao; Sun, Hongyang; Liang, Yi; Jiang, Haixiu; Song, Zhiling; Wang, Hong; Chen, Shizhong

    2016-02-01

    To study the interactions between natural compounds and deoxyribonucleic acid (DNA), a method has been established combining a high-performance liquid chromatography-diode array detector-multi-stage mass spectrometer with a fluorescence detector (HPLC-DAD-MS(n)-FLD). The FLD was used to monitor fluorescence intensity of the ethidium bromide-DNA (EB-DNA) complex when a compound separated by HPLC was introduced. This novel method was used to simultaneously obtain the HPLC fingerprint, UV spectra, MS(n) fragments and DNA-binding activity profile of various components in Folium Citri Reticulatae. As a result, 35 compounds were identified, of which 25 were found in the extract of Folium Citri Reticulatae for the first time, and 33 compounds showed DNA-binding activities, with the most active being feruloylhexaric and p-coumaroylhexaric acids. In addition, the precision, stability and reproducibility of this method were validated by two positive controls, quercetin and hesperidin. This new on-line method is accurate, precise and reliable for further high-throughput screening of DNA-binding compounds from food samples and other complex matrices.

  5. Study on Infrared Fingerprint of Exocarpium Citri Grandis with Different Species%化橘红药材红外指纹图谱的研究

    Institute of Scientific and Technical Information of China (English)

    卢丽萍; 林励; 黄爱华; 欧立娟; 汪金玉

    2011-01-01

    Objective To provide theoretical basis and practical methods for the rapid identification of Exocarpium Citri Grandis (ECG) with different species. Methods The fingerprints were established by fourier transform ifrared spectroscopy(FTIR), and then were analyzed by Spectrum v5.0.1 software. Results With normalization of the selected points between 1,820 cm-1 and 950 cm-1 on the FTIR fingerprints of ECG powder, significant differences were shown, and the second derivative spectra of the two different species between 1,800 cm-1 and 700 cm-1 had significant differences, which can be taken as one of the major identical characteristics to distinguish the two species. Conclusion The FTIR fingerprints is effective and feasible for the identification of the two species of ECG.%目的 建立化橘红药材的红外指纹图谱,通过考察不同品种化橘红药材的红外指纹图谱差异,为化橘红药材的快速鉴别提供理论依据和实用方法.方法 采用傅里叶红外光谱法(FTIR)建立不同品种化橘红药材红外指纹图谱,应用Spectrum v5.0.1红外软件对实验数据进行处理.结果 FTIR图谱在1820~950 cm-1范围内选点归一化处理后毛橘红和光橘红差异显著,其在1800~700 cm-1范围内二阶导数谱差异亦显著,可作为两者主要鉴别特征,经处理后的FTIR指纹图谱能有效鉴别两类化橘红药材.结论 红外指纹图谱法鉴别化橘红快速、简便、可行,可有效区分毛橘红和光橘红.

  6. Aspectos biológicos da fase adulta de Chrysoperla externa (Hagen, 1861 (Neuroptera: Chrysopidae oriunda de larvas alimentadas com Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae Biological aspects of the adult stage of Chrysoperla externa (Hagen, 1861 (Neuroptera: Chrysopidae originating from the larvae fed Planococcus citri (Risso, 1813 (Hemiptera: Pseudococcidae

    Directory of Open Access Journals (Sweden)

    Gerane Celly Dias Bezerra

    2006-08-01

    Full Text Available Objetivou-se estudar alguns aspectos biológicos da fase adulta de Chrysoperla externa (Hagen quando as larvas desse predador foram alimentadas com ninfas e fêmeas adultas de Planococcus citri (Risso. O experimento foi conduzido em câmara climatizada regulada a 25 ± 1 ºC, 70 ± 10% de UR e fotofase de 12 horas. Os adultos de C. externa cujas larvas foram alimentadas com ninfas nos três ínstares e fêmeas adultas da cochonilha, foram separados por sexo e 25 casais foram individualizados em gaiolas determinando-se a razão sexual, período de pré-oviposição, capacidade de oviposição diária e total durante 40 dias consecutivos, longevidade, período embrionário e viabilidade de ovos. Constatou-se que as ninfas nos três ínstares e fêmeas adultas fornecidas como alimento, não afetaram a razão sexual, período de pré-oviposição, período embrionário e longevidade dos adultos de C. externa. A produção diária e total de ovos por C. externa foi afetada pelo tipo de alimento fornecido ao predador durante a fase de larva, observando-se uma maior produção quando os casais foram alimentados na fase de larva com fêmeas adultas da cochonilha. Analisando-se a produção de ovos, observou-se uma maior concentração de oviposição nos primeiros dez dias do período reprodutivo, decrescendo nas avaliações seguintes. Foram obtidas porcentagens maiores de ovos viáveis e menores de inviáveis e inférteis, porém, não diferiram em função do alimento fornecido ao predador na fase de larva.It was aimed to study some biological aspects in the adult stage of Chrysoperla externa (Hagen when the larvae of predator were fed both nymphs and adult females of Planococcus citri (Risso. The experiment was conducted in climate chamber at 25 ± 1°C, 70 ± 10% of RH and 12-h photophase. The adults of C. externa whose larvae were fed nymphs at the three instars and adult females of mealybug were separated by sex and 25 couples were individualized

  7. Amostragem, caracterização de sintomas e escala diagramática da mancha graxa dos citros (Mycosphaerella citri no Recôncavo Baiano Sampling, characterization of citrus greasy spot (Mycosphaerella citri symptoms in Recôncavo Baiano and development of a diagrammatic scale for assessment of severity

    Directory of Open Access Journals (Sweden)

    Suely Xavier de Brito Silva

    2009-06-01

    Full Text Available O Recôncavo da Bahia é uma região de clima propício à mancha graxa dos citros (Mycosphaerella citri, doença nunca antes estudada no Brasil. Os objetivos deste trabalho foram definir o tamanho mínimo de amostra para quantificar a incidência, caracterizar os sintomas da doença e elaborar uma escala diagramática para a avaliação de sua severidade. A partir de uma amostragem piloto realizada por avaliação de cinco folhas por quadrante, quatro quadrantes por planta e 30 plantas em cada um dos 10 pomares visitados no município de Cruz das Almas, foi determinado que o tamanho mínimo da amostra em quadrantes e folhas seria de quatro e 16 plantas, respectivamente. De 320 folhas coletadas de diferentes plantas em 11 pomares, foi constatada uma grande quantidade de lesões (média de 131 por folha sintomática, em sua maioria diminutas (média de 0,014cm². A severidade variou de 0,15% a 35,85%, com média de 7,3%. Com base na severidade real em campo, foi elaborada uma escala diagramática com seis níveis: 1%, 2%, 5%, 9%, 18% e 36% de área foliar lesionada. Cinqüenta imagens de folhas foram submetidas a avaliadores por duas vezes, os quais estimaram a severidade com e sem a utilização da escala, constatando-se um discreto benefício no grau de precisão e de acurácia das estimativas com o uso da escala.Weather conditions make the Recôncavo Region in the State of Bahia, a favorable region to the occurrence of citrus greasy spot (Mycosphaerella citri, a disease that hadn't been studied before in Brazil. Thus, this research aimed to develop a sampling method to quantify its incidence, to characterize its symptoms, and to develop a diagrammatic scale for assessing disease severity. The minimum sample size to estimate greasy spot incidence in leaves (16 plants was calculated based on a prospective sampling of five leaves per quadrant, four quadrants per tree, and 30 trees per grove, in ten groves. From 320 mature leaves collected in

  8. Investigation on the resistance of different varieties of citrus to Xanthomonas citri%不同柑橘品种对柑橘溃疡病的抗性调查

    Institute of Scientific and Technical Information of China (English)

    宋盛英; 杨光兰; 潘盛勇; 赵春穆; 石安鸠

    2002-01-01

    通过柑橘Citrus sp.4类10个品种对柑橘溃疡病Xanthomonas citri(Hasse)Dowson抗病性的调查发现,不同的柑橘品种对柑橘溃疡病的抗病程度有明显差异,甜橙类、脐橙类普遍容易感病,属高度感病品种;宽皮橘类的抗病性最强;而柚类感病轻重不一,以台湾白柚抗病性最强,而沙田柚抗病性较弱.

  9. 柑橘全爪螨微卫星位点鉴定与信息分析%Analysis of Microsatellite Loci fromPanonychus citri Based on Enriched Microsatellite Library and Transcriptome Dataset

    Institute of Scientific and Technical Information of China (English)

    魏丹丹; 刘燕; 杜洋; 李刚; 李婷; 袁明龙; 王进军

    2016-01-01

    重复类型,占总数的53.86%;其次为两碱基核心重复类型,占总数的43.36%;而四、五、六碱基重复类型以及复合型的SSR数量均较少,且数量差异不大,共占EST-SSR总数的2.78%。柑橘全爪螨EST-SSR核心重复次数主要集中在5—10次。【结论】采用磁珠富集法,并将酶切、接头连接一步化,可提高个体微小的螨类及微小昆虫SSR的富集效率。柑橘全爪螨gSSR核心重复次数要远多于从转录组数据库获得的EST-SSR。总体而言,gSSR和EST-SSR中的三碱基重复SSR具有更好的优化率。此外,柑橘全爪螨gSSR具有微卫星家族现象。%Objective] The objective of this study is to construct the microsatellite-enriched libraries, and identify genomic- microsatellite (gSSR) from the genome of the citrus red mite,Panonychus citri. Meanwhile, a large number of gene-microsatellite (EST-SSR) markers were also indentified from the transcriptome database ofP. citri. Based on these SSR sequences, the selected SSR primer pairs were validated.[Method]On the basis of isolating high quality genomic DNA ofP. citri, enriched microsatellite libraries were constructed by streptavidin-coated magnetic beads utilizing methodologies that exploit the strong affinity between biotin and the protein streptavidin. A fast and easy protocol was proposed through a combination of two different published methods. Briefly, genomic DNA was digested by the restriction enzyme and then ligated to designed adaptors. Microsatellite-containing DNA fragments were captured by streptavidin-coated magnetic beads. The beads affinity capture of microsatellite repeats using biotinylated oligonucleotide probes. Subsequently, PCR was used to amplify the captured molecules for transferring single strand DNA to double strand DNA. The PCR products were then ligated to pGEM-T Easy vector and transformed into Trans5α competent cells. Then, clones from these libraries were screened for microsatellite content. Meanwhile

  10. The GC-MS analysis of volatile oil from Atractylodes Macrocephala Koidz,Pericarpium Citri Reticulate and their mixed samples%药对白术-陈皮挥发油成分GC-MS分析

    Institute of Scientific and Technical Information of China (English)

    杨小红; 罗琼; 胡剑卓; 陈文; 谭敏

    2012-01-01

    Objective To analysis the volatile components in herbal pair Atractylodes Macrocephala -Pericarpium Citri Reticulatae (HP AM-PCR), single herb Atractylodes Macrocephala(AM) and Pericarpium Citri Reticulatae (PCR) quantitatively. Methods Volatile oil from above materials were obtained with water reflux condensation method;oil composition was detected by gas - mass spectrometry (GC-MS) and the data was analyzed with heuristic evolving latent projection (HELP); Comparing the difference between chromatographys to identify the main chemical constituents. Results There was no new compound found in herbal pair. The compounds were from single herbs. Conclusion Chemical changes in herbal pair AM -PCR were physical compatibility.%目的:定性、定量分析药对白术-陈皮、单味药白术和陈皮的主要挥发油成分,探讨药对和单味药挥发油成分变化.方法:共水回流冷凝法分别提取单味白术、陈皮及药对挥发油、气相-质谱(GC-MS)检测,直观推导式演进特征投影(HEIP)法解析所得GC-MS数据,比较图谱差异,鉴别主要化学成分.结果:药对白术-陈皮主要组分中未发现新的结构类型化合物,挥发油化合物来源于单味药材,且主要为白术挥发油.结论:药对白术-陈皮挥发油化学成分变化属于物理配伍变化.

  11. Annotation of the Asian Citrus Psyllid Genome Reveals a Reduced Innate Immune System.

    Science.gov (United States)

    Arp, Alex P; Hunter, Wayne B; Pelz-Stelinski, Kirsten S

    2016-01-01

    Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as Huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Hemipterans characterized to date lack a number of insect immune genes, including those associated with the Imd pathway targeting Gram-negative bacteria. The D. citri draft genome was used to characterize the immune defense genes present in D. citri. Predicted mRNAs identified by screening the published D. citri annotated draft genome were manually searched using a custom database of immune genes from previously annotated insect genomes. Toll and JAK/STAT pathways, general defense genes Dual oxidase, Nitric oxide synthase, prophenoloxidase, and cellular immune defense genes were present in D. citri. In contrast, D. citri lacked genes for the Imd pathway, most antimicrobial peptides, 1,3-β-glucan recognition proteins (GNBPs), and complete peptidoglycan recognition proteins. These data suggest that D. citri has a reduced immune capability similar to that observed in A. pisum, P. humanus, and R. prolixus. The absence of immune system genes from the D. citri genome may facilitate CLas infections, and is possibly compensated for by their relationship with their microbial endosymbionts.

  12. The controlling role of Panonychus citri by "Lümanyuan" in the filed and the measurement of its acute toxicity to Carassius auratus%“绿满园”对柑桔红蜘蛛的田间防治效果及对金鱼的毒性测定

    Institute of Scientific and Technical Information of China (English)

    乔虎平; 王金亮; 江庆红; 云月利; 王维; 彭宇

    2013-01-01

    测定“绿满园”水剂对柑桔红蜘蛛Panonychus citri的田间防治效果及对非靶标生物金鱼Carassius auratus 的急性毒性.结果表明,64%“绿满园”水剂800倍和1 000倍稀释液在施药后7d和15d对柑桔红蜘蛛的防治效果均极显著地高于对照杀螨剂:15%扫螨净和57%炔螨特,其中以800倍稀释液防治效果最好,15d的防效达97.21%,800倍和1 000倍稀释液的防治效果差异不明显.金鱼的死亡率随“绿满园”浓度的升高和染毒时间的延长逐渐增加.“绿满园”水剂对金鱼的急性毒性小于阿维菌素.%The controlling role of Panonychus citri by "Lumanyuan" in the filed and the measurement of its acute toxicity to Carassius auratus were carried out. The results showed that, 15 days after spraying, the controlling roles of 800 folds and 1 000 folds dilution of 64% "Lumanyuan" to P. citri was obviously higher than 15% pyridaben and 57% propargite. 800 fold dilution of "Lumanyuan" had the best controlling role to P. citri and the controlling effect was 97. 21%. There was no significant difference on controlling effects between 800 folds and 1 000 folds dilution of "Lumanyuan" to P. citri. The mortalities of C. auratus increased with the rise of the concentration and treatment time of "Lumanyuan" and "Lumanyuan" has lower acute toxicity to C. auratus than avermectin.

  13. Regular LAMP and fast LAMP for the detection of Xanthomonas axonopodis pv.citri%柑橘溃疡病菌的普通LAMP及快速LAMP检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    张仑; 殷幼平; 吴瑜佳; 王中康

    2013-01-01

    This study is aimed to establish the regular LAMP and fast LAMP for the detection of Xanthomonas axonopodis pv.citri and make it useful for the grassroots quarantine departments.A specific region of the genome of X.axonopodis pv.citri was used for the design of regular LAMP and fast LAMP primers; the regular LAMP and fast LAMP detection system were established by optimization of reaction conditions; the specificity of the system was verified using genomic DNA of several reference strains and healthy citrus leaves by using a series of gradient dilution of DNA solution of X.axonopodis pv.citri and bacterial suspension.Bacterial suspension and DNA sensitivity of regular LAMP system reached 2.25 × 104 cfu and 2.03× 10-1 ng,respectively,while those of fast LAMP system reached 2.25 cfu and 2.03× 10-5 ng,respectively.In the specificity test,both regular LAMP and fast LAMP system showed high and identical specificity.The total time for the fast LAMP reaction was within 30 minutes,only half of that for the regular LAMP,greatly improving the efficiency of the analysis.The fast LAMP system was ten thousand times as sensitive as the regular LAMP system.Regular LAMP and fast LAMP for the detection of X.axonopodis pv.citri was successfully established.Regular LAMP and fast LAMP system provide a new,fast and easy way for the detection of X.axonopodis pv.citri.%建立柑橘溃疡病菌的普通LAMP和快速LAMP检测方法,使其能应用于基层检验检疫部门对病害的快速检测.利用柑橘溃疡病菌基因组特有的保守区域设计LAMP引物,通过优化反应条件,建立柑橘溃疡病菌的普通LAMP检测体系;在普通LAMP引物的基础上设计一对环引物,建立柑橘溃疡病菌的快速LAMP检测体系,并以多种参比菌DNA以及健康柑橘叶片基因组DNA为模板对普通LAMP和快速LAMP检测体系的特异性进行了验证,利用柑橘溃疡病菌菌液和DNA溶液梯度稀释液对普通LAMP和快速LAMP检测体系的灵敏度进

  14. 阿维菌素与印楝油对柑桔红蜘蛛的增效作用%Synergistic Effect of Abamectin and Neem Oil Against Panonychus citri Me Gregor

    Institute of Scientific and Technical Information of China (English)

    熊忠华; 李保同; 熊件妹; 张纪利

    2012-01-01

    Toxicity of abamectin and neem oil against Panonychus citri and the synergistic effect of their mixture were evaluated in the lab. It was showed that, for the contact toxicity of abamectin and neem oil to adult famales of Panonychus citri, the LCX value was 0.01 jig/mL and 6 579.87 u.g/mL, respectively, after 24 h treatment. The optimal mixture ratio of abamectin with neem oil was determined by co-toxicity factor and eo-toxicity coefficient method. The results indicated that, among the 6 combinations of theses two active ingredients, the co-toxicity factor of combination at the ratios of 1 : 100 and 1 : 125 for abamectin vs neem oil was 24.50 and 31.08, respectively. Furthermore, combination at the ratio of 1 : 125 showed the highest synergism with a co-toxicity coefficient of 160.76.%室内测定阿维菌素与印楝油对柑桔红蜘蛛的毒力及其复配组合的联合作用.毒力测定结果表明,阿维菌素与印楝油对柑桔红蜘蛛雌成螨处理后24 h的LC50分别为0.01 μg/mL和6579.87 μg/mL.以共毒因子法评价阿维菌素与印楝油6种复配组合的联合作用,结果显示,二者以1∶100和1∶125两种比例复配对柑桔红蜘蛛雌成螨的共毒因子分别为24.50和31.08,具增效作用;进一步以共毒系数法确定阿维菌素与印楝油以1∶125的比例复配增效作用最为显著,共毒系数为160.76.

  15. Asian Citrus Psyllid RNAi Pathway – RNAi evidence

    OpenAIRE

    Taning, Clauvis N. T.; Eduardo C. Andrade; Hunter, Wayne B.; Olivier Christiaens; Guy Smagghe

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if t...

  16. Asian Citrus Psyllid RNAi Pathway – RNAi evidence

    OpenAIRE

    Taning, Clauvis N. T.; Andrade, Eduardo C. de; Hunter, Wayne B.; Christiaens, Olivier; Smagghe, Guy

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if t...

  17. Asian citrus psyllid RNAi pathway : RNAi evidence

    OpenAIRE

    Taning, Clauvis N. T.; Andrade, Eduardo C.; Hunter, Wayne B.; Olivier Christiaens; Guy Smagghe

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if t...

  18. 防治橘臀纹粉蚧药剂的室内筛选%Insecticide Screening for the control of Planococcus citri(Risso)(Homoptera: Pseudococcidae) in the laboratory

    Institute of Scientific and Technical Information of China (English)

    郭莹; 刘长明; 吴梅香; 杨广

    2012-01-01

    为有效地控制橘臀纹粉蚧Planococcus citri(Risso)发生,测定了6种农药在田间使用浓度下对该虫的致死效果.结果表明,在室内条件下,吡虫啉、烯啶虫胺、啶虫脒、阿维菌素和高效氯氰菊酯对1龄若虫的防治效果较好,用药后72 h校正死亡率均在80.0%以上,而杀扑磷仅74.8%.在防治应用中,建议推荐吡虫啉,烯啶虫胺,啶虫脒3种药剂,同时应针对不同虫态适当调整用药浓度和施药时间,以达到理想的防治效果.

  19. Synergistic Effect of Three Substances on Paris polyphylla Smith against Panoychus citri McGregor%3种物质对七叶一枝花防治柑桔全爪螨的增效作用

    Institute of Scientific and Technical Information of China (English)

    周顺玉; 蒋春先; 杨群芳; 李庆

    2007-01-01

    研究了邻苯二酚、氮酮和硫酸钙3种化学物质对七叶一枝花(Paris polyphylla Smith)提取物防治柑桔全爪螨(Panoychus citri McGregor)的增效作用.结果表明:七叶一枝花乙醇、氯仿、乙酸乙酯提取物单用对柑桔全爪螨的致死中浓度(LC50)分别为1 588.537、1480.071、1 402.954 μg/ml;邻苯二酚对七叶一枝花乙醇、氯仿、乙酸乙酯提取物均有显著增效作用,LC50分别为284.506、582.295、897.323 μg/ml,分别增效5.6、2.5和1.6倍;氮酮仅对七叶一枝花乙醇提取物有增效作用,增效2.1倍;硫酸钙对七叶一枝花氯仿、乙酸乙酯提取物均有增效作用,分别增效1.4和2.1倍.

  20. Microwave-based Method for Genomic DNA Extraction and Rapid Detection Method from Xanthomonas axonopodis pv.citri%柑桔溃疡病菌基因组DNA的微波法抽提及快速检测

    Institute of Scientific and Technical Information of China (English)

    姚廷山; 周常勇; 胡军华; 冉春; 李鸿筠; 刘浩强; 肖田

    2012-01-01

    利用微波热震惊提取固体表面柑桔溃疡病基因组DNA并加以优化,所得到的基因组DNA可作为PCR反应的模板进行16SrDNA基因有效扩增.与柑桔溃疡病基园组DNA其他抽提方法相比较,微波法更适用于该病的快速检测,具有快速、筒便、费用低廉等特点,且对设备的要求不高,用特异性引物XCF/XCR可实现柑桔溃疡病菌的快速鉴定.%The optimized microwave extraction was conducted to attract the genome of citrus canker disease. The genome extraction method was fast,easy to handle with low cost The extracted DNA was suitable for PCR. The microwave-based method was suitable for fast identification compare to other methods. The method was effective, easy and fast, so it was recommended that the extraction method was applied for the detection of Xanthomonas oxonopodit pv. citri using primers XCF/XCR.

  1. Investigation on Chelidonium citri attacking Murraya paniculata and its control%橘光绿天牛危害九里香绿化球调查及防治

    Institute of Scientific and Technical Information of China (English)

    刘有莲; 黄寿昌

    2003-01-01

    九里香绿化球受橘光绿天牛Chelidonium citri Grressitt为害3~4 a后,受害株率90.7%,枝枯株率74.1%,虫口密度2.11,危害度31.5%.危害造成枝枯形成空洞平均宽度为61 cm,占树冠平均周长的14.4%;枝枯形成空洞平均深度为68.4 cm,占平均树高50.8%.植株受害与方向的关系为:植株东向受害最多,占受害总株数30%;中部枝枯形成空洞平均宽度占树冠平均周长比率最大,为22.9%;西南方向枝枯形成空洞平均深度最深,占平均高度的90.9%.植株周围阴湿,虫口密度大.

  2. A specific interdomain interaction preserves the structural and binding properties of the ModA protein from the phytopathogen Xanthomonas citri domain interaction and transport in ModA.

    Science.gov (United States)

    Santacruz-Perez, Carolina; Pegos, Vanessa Rodrigues; Honorato, Rodrigo V; Verli, Hugo; Lindahl, Erik; Barbosa, João Alexandre Ribeiro Gonçalves; Balan, Andrea

    2013-11-01

    The periplasmic-binding proteins in ATP-binding cassette systems (ABC Transporters) are responsible for the capture and delivery of ligands to their specific transporters, triggering a series of ATP-driven conformational changes that leads to the transport of the ligand. Structurally consisting of two lobes, the proteins change conformation after interaction with the ligand. The structure of the molybdate-binding protein (ModA) from Xanthomonas citri, bound to molybdate, was previously solved by our group and an interdomain interaction, mediated by a salt bridge between K127 and D59, apparently supports the binding properties and keeps the domains closed. To determinate the importance of this interaction, we built two ModA mutants, K127S and D59A, and analysed their functional and structural properties. Based on a set of spectroscopic experiments, crystallisation trials, structure determination and molecular dynamics (MD) simulations, we showed that the salt bridge is essential to maintain the structure and binding properties. Additionally, the MD simulations revealed that this mutant adopted a more compact structure that packed down the ligand-binding pocket. From the closed bound to open structure, the positioning of the helices forming the dipole and the salt bridge are essential to induce an intermediate state. Copyright © 2013 Elsevier Inc. All rights reserved.

  3. Harm of Chelidonium citri Grressitt to Murraya paniculata (L.) Jack Balls and Economic Damage Appraisal%橘光绿天牛对九里香绿化球的危害及其经济损失评估

    Institute of Scientific and Technical Information of China (English)

    刘有莲; 黄寿昌

    2003-01-01

    分别于2002年6月27日和8月29日调查广西生态工程职业技术学院内种植的九里香[Murrayapaniculata(L.)Jack]受橘光绿天牛(Chelidomum citri Grressitt)危害的状况并进行危害价值损失评估.结果表明,九里香绿化球受害株率90.7%,枝枯株率74.1%,有虫株率75.9%,虫口密度2.11,危害度31.5%.危害造成枝枯宽度平均为61 cm,占平均树冠周长的14.4%;枝枯平均深度为68.4cm,占平均树高50.8%.植株东向受害最多,占受害总株数30%;中部枝枯宽度占树冠周长比例最大,为22.9%;西南方向枝枯深度最深,占高度的90.9%.九里香绿化球价值损失占总价值的38.7%.学院的九里香绿化球受橘光绿天牛危害严重,需要及时有效的预防与防治.

  4. A plant pathogenic bacterium exploits the tricarboxylic acid cycle metabolic pathway of its insect vector.

    Science.gov (United States)

    Killiny, Nabil; Nehela, Yasser; Hijaz, Faraj; Vincent, Christopher I

    2017-06-08

    Huanglongbing in citrus is caused by a phloem-limited, uncultivable, gram-negative α-proteobacterium, Candidatus Liberibacter asiaticus (CLas). CLas is transmitted by the phloem-sucking insect, Diaphorina citri (Hemiptera: Liviidae), in a persistent, circulative, and propagative manner. In this study, we investigated the metabolomic and respiration rates changes in D. citri upon infection with CLas using gas chromatography-mass spectrometry (GC-MS) and gas exchange analysis. The level of glycine, L-serine, L-threonine, and gamma-amino butyric acid were higher in CLas-infected D. citri, while L-proline, L-aspartic acid, and L-pyroglutamic acid were lower in CLas-infected D. citri compared with the control. Citric acid was increased in CLas-infected D. citri, whereas malic and succinic acids were reduced. Interestingly, most of the reduced metabolites such as malate, succinate, aspartate, and L-proline are required for the growth of CLas. The increase in citric acid, serine, and glycine indicated that CLas induced glycolysis and the tricarboxylic acid cycle (TCA) in its vector. In agreement with the GC-MS results, the gene expression results also indicated that glycolysis and TCA were induced in CLas-infected D. citri and this was accompanied with an increases in respiration rate. Phosphoric acid and most of the sugar alcohols were higher in CLas-infected D. citri, indicating a response to the biotic stress or cell damage. Only slight increases in the levels of few sugars were observed in CLas-infected D. citri, which indicated that sugars are tightly regulated by D. citri. Our results indicated that CLas induces nutrient and energetic stress in its host insect. This study may provide some insights into the mechanism of colonization of CLas in its vector.

  5. Extracts of Immature Orange (Aurantii fructus immaturus) and Citrus Unshiu Peel (Citri unshiu pericarpium) Induce P-Glycoprotein and Cytochrome P450 3A4 Expression via Upregulation of Pregnane X Receptor.

    Science.gov (United States)

    Okada, Naoto; Murakami, Aki; Urushizaki, Shiori; Matsuda, Misa; Kawazoe, Kazuyoshi; Ishizawa, Keisuke

    2017-01-01

    P-glycoprotein (P-gp) and cytochrome P450 3A4 (CYP3A4) are expressed in the intestine and are associated with drug absorption and metabolism. Pregnane X receptor (PXR) is the key molecule that regulates the expression of P-gp and CYP3A4. Given that PXR activity is regulated by a variety of compounds, it is possible that unknown PXR activators exist among known medicines. Kampo is a Japanese traditional medicine composed of various natural compounds. In particular, immature orange [Aurantii fructus immaturus (IO)] and citrus unshiu peel [Citri unshiu pericarpium (CP)] are common ingredients of kampo. A previous study reported that kampo containing IO or CP decreased the blood concentration of concomitant drugs via upregulation of CYP3A4 although the mechanism was unclear. Some flavonoids are indicated to alter P-gp and CYP3A4 activity via changes in PXR activity. Because IO and CP include various flavonoids, we speculated that the activity of P-gp and CYP3A4 in the intestine may be altered via changes in PXR activity when IO or CP is administered. We tested this hypothesis by using LS180 intestinal epithelial cells. The ethanol extract of IO contained narirutin and naringin, and that of CP contained narirutin and hesperidin. Ethanol extracts of IO and CP induced P-gp, CYP3A4, and PXR expression. The increase of P-gp and CYP3A4 expression by the IO and CP ethanol extracts was inhibited by ketoconazole, an inhibitor of PXR activation. The ethanol extract of IO and CP decreased the intracellular concentration of digoxin, a P-gp substrate, and this decrease was inhibited by cyclosporine A, a P-gp inhibitor. In contrast, CP, but not IO, stimulated the metabolism of testosterone, a CYP3A4 substrate, and this was inhibited by a CYP3A4 inhibitor. These findings indicate that the ethanol extract of IO and CP increased P-gp and CYP3A4 expression via induction of PXR protein. Moreover, this induction decreased the intracellular substrate concentration.

  6. Extracts of Immature Orange (Aurantii fructus immaturus) and Citrus Unshiu Peel (Citri unshiu pericarpium) Induce P-Glycoprotein and Cytochrome P450 3A4 Expression via Upregulation of Pregnane X Receptor

    Science.gov (United States)

    Okada, Naoto; Murakami, Aki; Urushizaki, Shiori; Matsuda, Misa; Kawazoe, Kazuyoshi; Ishizawa, Keisuke

    2017-01-01

    P-glycoprotein (P-gp) and cytochrome P450 3A4 (CYP3A4) are expressed in the intestine and are associated with drug absorption and metabolism. Pregnane X receptor (PXR) is the key molecule that regulates the expression of P-gp and CYP3A4. Given that PXR activity is regulated by a variety of compounds, it is possible that unknown PXR activators exist among known medicines. Kampo is a Japanese traditional medicine composed of various natural compounds. In particular, immature orange [Aurantii fructus immaturus (IO)] and citrus unshiu peel [Citri unshiu pericarpium (CP)] are common ingredients of kampo. A previous study reported that kampo containing IO or CP decreased the blood concentration of concomitant drugs via upregulation of CYP3A4 although the mechanism was unclear. Some flavonoids are indicated to alter P-gp and CYP3A4 activity via changes in PXR activity. Because IO and CP include various flavonoids, we speculated that the activity of P-gp and CYP3A4 in the intestine may be altered via changes in PXR activity when IO or CP is administered. We tested this hypothesis by using LS180 intestinal epithelial cells. The ethanol extract of IO contained narirutin and naringin, and that of CP contained narirutin and hesperidin. Ethanol extracts of IO and CP induced P-gp, CYP3A4, and PXR expression. The increase of P-gp and CYP3A4 expression by the IO and CP ethanol extracts was inhibited by ketoconazole, an inhibitor of PXR activation. The ethanol extract of IO and CP decreased the intracellular concentration of digoxin, a P-gp substrate, and this decrease was inhibited by cyclosporine A, a P-gp inhibitor. In contrast, CP, but not IO, stimulated the metabolism of testosterone, a CYP3A4 substrate, and this was inhibited by a CYP3A4 inhibitor. These findings indicate that the ethanol extract of IO and CP increased P-gp and CYP3A4 expression via induction of PXR protein. Moreover, this induction decreased the intracellular substrate concentration. PMID:28270768

  7. Application of immunoaffinity purification technology as the pretreatment technology for traditional Chinese medicine: Its application to analysis of hesperidin and narirutin in traditional Chinese medicine preparations containing Citri reticulatae Pericarpium.

    Science.gov (United States)

    Liu, Li-Na; Wang, Ying; Jin, Hong-Yu; Ma, Shuang-Cheng; Liu, Jia-Peng

    2016-05-15

    In the present study, the feasibility of immunoaffinity chromatography (IAC) as a purification technology for the analysis of bioactive components in Traditional Chinese Medicine (TCM) was evaluated. IAC was used to analyze hesperidin (HP) and narirutin (NR) in TCM preparations containing Citri reticulatae Pericarpium (CRP, Chenpi in Chinese). An IAC column for the specific extraction and enrichment of HP and NR from TCM preparations containing CRP was developed and characterized. After HP reacted with carbonyl diimidazole and coupled to protein, it was used to immune mice for the generation of antibody. Through cell fusion, cloning and screening, monoclonal antibody was obtained. The IAC column was constructed by covalently coupling specific monoclonal antibody against HP and NR to CNBr-activated Sepharose 4B and packed into a common solid phase extraction cartridge. The extraction conditions including loading, washing and eluting, as well as flow rate for the extraction of HP and NR were optimized. Under optimal conditions, the maximum capacity, extraction recovery rate and stability of IAC column was also characterized. Results revealed that the maximum capacity of IAC column for HP and NR was approximately 16μg and the relative binding capacity per 1mL of the column volume was 27μg. The extraction recovery rate of IAC column for HP and NR at three spiked levels was in the range of 94.05-109.15%. After the repeated application for 5 times, no significant loss of specific recognition was observed. Using high performance liquid chromatography (HPLC) as an effective analytic tool, HP and NR could be successfully separated via IAC column without the inference from impurities, suggesting that the extraction of HP and NR using the prepared IAC column is feasible. The application of IAC can solve the problem of quantitative analysis due to severe interference or low content. Furthermore, pretreatment methods in different matrixes can be unified. The IAC purification

  8. Study on the Dynamic Change Law of Essential Oil Components in Citri Reticulatae Pericarpium from the Same Plant%源于同一植株陈皮挥发油成分动态变化规律研究

    Institute of Scientific and Technical Information of China (English)

    王坚; 韦正; 陈鸿平; 银玲; 刘荣; 刘友平

    2013-01-01

    目的:研究源于同一植株不同生长期陈皮挥发油成分的变化规律.方法采用水蒸气蒸馏法提取挥发油,GC-MS测定;GC-MS数据分析与AMDIS软件进行解析.结果:4批陈皮共定性、定量72个化合物,总含量分别为95.542%,92.138%,98.655%,96.201%.含量最高的2个化合物是柠檬烯与芳樟醇,分别占43.78 ~ 68.432%,9.258 ~ 15.724%.结论:4批次陈皮挥发油几乎含有相同的物质成分,各成分在含量上有一定差异.一些物质的量变规律对于揭示陈皮挥发油类成分的动态变化规律具有重要意义.%Objective: To investigate the qualitative and quantitative change law of essential oil components in Citri Reticulatae Pericarpium (CRP) harvested in different time from the same plant. Method: The volatile oils were extracted by steam distillation and analyzed by GC-MS. The overlapped peaks were resolved by the programs of GC-MS data analysis and AMDIS. Result: A total of 72 compounds in the essential oils of four batches CRP were identified, which represented 95.542% , 92. 138% , 98.655% , and 96.201% of the total relative content, respectively. The two compounds that have the highest content are limonene and linalool, accounting for 43.78-68.432% and 9.258-15. 724% respectively. Quantitative changes of some compounds present regularity. Conclusion: The essential oil of each batch CRP has almost the same chemical constituents, whereas the relative content of each constituent shows some difference. The quantitative change law of some components may have important meaning to explore the dynamic change law of volatile oil components in CRP. The combined utilization of programs of GC-MS data analysis and AMDIS can efficiently solve the difficulty in the qualitative and quantitative analysis of overlapped peaks.

  9. Screening Experiments for the Effective Fungicides of Black Rot Disease of Gonggan(Alternaria citri)%贡柑黑腐病有效防治药剂筛选

    Institute of Scientific and Technical Information of China (English)

    邓明学; 谢艳芳; 谭有龙; 覃旭; 覃星瑜; 覃宗典

    2012-01-01

    [目的]筛选防治贡柑黑腐病的有效药剂.[方法]用生长速率法和孢子萌发法测定苯醚甲环唑等15种药剂的EC50、EC95值,并做氟硅唑等5种药剂的田间药效试验.[结果]以EC95(mg a.i./L)排序,抑菌活性较高的有氟硅唑、吡唑醚菌酯·咪鲜胺、吡唑醚菌酯、苯醚甲环唑、咪鲜胺、代森锰锌、丙森锌、百菌清·多菌灵·代森锰锌;田间防效较好的有10%氟硅唑WP、25%苯醚甲环唑·咪鲜胺WP、50%百菌清·多菌灵·代森锰锌WP和72%代森锰锌·霜脲氰WP,而10%氟硅唑EW防效较差.[结论]耐雨水冲刷的杀菌剂和剂型防效较好.%[Aims] This paper aims to screen out the effective fungicides for controlling black rot disease of Gonggan(Alteranria citri). [Methods] The EC50 and EC95(mg a.i./L) of 15 different kinds of fungicides, such as ifenoconazole, pyraclostrobin, were tested with the both methods of the growth rate and spore emergence, meanwhile, the field control of five kinds of fungicides, such as flusilazole, were conducted. [Results] Sorting by EC,,, the effect on inhibition of mycelial growth and spore germination from high to low: flusilazole, pyraclostrobin+prochloraz, pyraclostrobin, difenoconazole, prochloraz, mancozeb, propineb. The field trial results showed that the following .fungicides have good effect: flusilazole 10% WP 1 g/L, difenoconazole+prochloraz 25% WP 1 g/L, chlorothalonil+carbendazim+mancozeb 50% WP 1.67 g/L and mancozeb+cymoxanil 72% WP 1.25 g/L, while flusilazole 10% EW 1 g/L with poor control efficiency. [Conclusions] The control effect is better of the fungicides whose formulations can resist rain erosion.

  10. Rapid characterization of 96 chemical constituents in Citri Reticulatae Folium (leaves of ‘Fuju’) using HPLC-DAD-ESI-MSn%HPLC-DAD-ESI-MSn技术对橘叶(福橘叶)中96个化学成分的快速分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    曹规划; 付庆荣; 张藏曼; 王弘; 陈世忠

    2016-01-01

    为了对橘叶(福橘叶)中的化学成分进行较为系统地研究,我们首次采用液质联用技术((HPLC-DAD-ESI-MSn)系统地分离和鉴定了橘叶中的化学成分.最终,我们在橘叶中解析了96个化学成分,包括:31个酚酸类化合物,4个黄酮苷元,6个黄酮单氧苷,10个黄酮双氧苷,5个黄酮单碳苷和5个黄酮双碳苷化合物,以及6个黄酮碳氧苷,5个3-羟基-3-甲基戊二酰基取代的黄酮苷,1个黄烷-3-醇和2个生物碱类化合物,并且初步鉴定出了21个多甲基黄酮类化合物(PMFs).在上述解析的96个化合物中,52个化合物为首次在橘叶中发现,15个化合物为首次在柑橘属植物中发现.同时,我们在橘叶中也发现了9个潜在的新化合物.%In order to systematically investigate the chemical constituents of Citri Reticulatae Folium (leaves of ‘Fuju’),an analytical method that included high-performance liquid chromatography,diode array detection,electrospray ionization,and ion-trap time-of-flight mass spectrometry (HPLC-DAD-ESI-MSn) was used to separate and identify the individual chemical components of Citri Reticulatae Folium.As a result,96 compounds were tentatively identified in this study:including 31 phenolic acids,4 flavonoid aglycones,6 flavonoid mono-O-glycosides,10 flavonoid-O-diglycosides,5 flavonoid mono-C-glycosides,5 flavonoid di-C-glycosides,6 flavonoid O,C-glycosides,5 (3-hydroxy-3-methylglutaryl) glycosyl flavonoids,1 flavan-3-ol,and 2 alkaloids.In addition,21 polymethoxy flavonoids (PMFs) were identified in this paper.Among these compounds,52 compounds,which were previously found in other Citrus plants,have been identified for the first time in Citri Reticulatae Folium.15 compounds have not been previously found in the Citrus genus were identified.Moreover,9 potentially new compounds have also been detected in this paper.This is the first report of the full characterization of chemical components of Citri Reticulatae Folium (leaves of

  11. Difference analysis of GABA receptor gene between different strains in the red mite, Panonychus citri (Acari: Tetranychidae) of citrus%橘全爪螨不同品系γ氨基丁酸受体基因差异性分析

    Institute of Scientific and Technical Information of China (English)

    张云飞; 李晓娇; 岳建苏; 江高飞; 冉春

    2012-01-01

    【目的】为明确γ氨基丁酸受体基因与橘全爪螨抗性的关系,【方法】通过BLAST搜索,对GABA受体基因进行鉴定:进一步通过序列比对和Sanger测序,对橘全爪螨敏感品系和噻螨酮抗性品系GABA受体基因SNPs进行分析和真实性验证;采用RPKM法对橘全爪螨敏感品系和噻螨酮抗性品系GABA受体基因进行表达差异分析。【结果】从橘全爪螨转录组中获得了19条GABA受体基因。通过敏感品系和抗性品系GABA受体基因序列比较和sanger测序发现Unigene11199_All有2个SNP位点。对GABA受体基因表达差异分析发现,相对于敏感品系,抗性品系中部分GABA受体基因表达量发生了不同程度的变化,抗性品系中有3条GABA受体基因表达上调,13条GABA受体基因表达下调,Unigene24440_All下调倍数最高[10g2Ratio(RS/SS)=-10.452479]。【结论】由此推断,橘全爪螨抗性产生可能是一个比较复杂的过程,而Unigenelll99_All和Unigene24440_All可能是橘全爪螨对噻螨酮产生抗性的重要原因。%[Objective]In order to clarify the relationship between T-aminobutyric acid receptor gene and resistance of Panonychus citri. [Method]GABA receptor genes were identified by BLAST searching. SNPs in GABA receptor genes between susceptible strain and hexythiazox-resistant strain of P. citri were ana- lyzed and examined by comparison of GABA receptor gene sequences and Sanger sequencing, and gene expression profiles of GABA receptor genes were further compared using RPKM (Reads Per kb per Million reads) method. [Result] 19 GABA receptor genes were found in transcripts of P. citri. Comparisons of GA- BA receptor gene sequences and further sequencing showed that there were 2 SNPs in Unigenel 1199_All between different strains. Analysis of difference of gene expression showed 3 GABA receptor genes were up-regulated and 13 were down-regulated, of which, Unigenel1069_All was the top down

  12. Metabolomic comparative analysis of the phloem sap of curry leaf tree (Bergera koenegii), orange jasmine (Murraya paniculata), and Valencia sweet orange (Citrus sinensis) supports their differential responses to Huanglongbing.

    Science.gov (United States)

    Killiny, Nabil

    2016-11-01

    Orange jasmine, Murraya paniculata and curry leaf tree, Bergera koenegii are alternative hosts for Diaphorina citri, the vector of Candidatus Liberibacter asiaticus (CLas), the pathogen of huanglongbing (HLB) in citrus. D. citri feeds on the phloem sap where CLas grows. It has been shown that orange jasmine was a better host than curry leaf tree to D. citri. In addition, CLas can infect orange jasmine but not curry leaf tree. Here, we compared the phloem sap composition of these 2 plants to the main host, Valencia sweet orange, Citrus sinensis. Phloem sap was analyzed by gas chromatography-mass spectrometry after trimethylsilyl derivatization. Orange jasmine was the highest in proteinogenic, non-proteinogenic amino acids, organic acids, as well as total metabolites. Valencia was the highest in mono- and disaccharides, and sugar alcohols. Curry leaf tree was the lowest in most of the metabolites as well as total metabolites. Interestingly, malic acid was high in Valencia and orange jasmine but was not detected in the curry leaf. On the other hand, tartaric acid which can prevent the formation of malic acid in Krebs cycle was high in curry leaf. The nutrient inadequacy of the phloem sap in curry leaf tree, especially the amino acids could be the reason behind the longer life cycle and the low survival of D. citri and the limitation of CLas growth on this host. Information obtained from this study may help in cultivation of CLas and development of artificial diet for rearing of D. citri.

  13. Insecticide resistance of adults and nymphs of Asian citrus psyllid populations from Apatzingán Valley, Mexico.

    Science.gov (United States)

    Pardo, Saúl; Martínez, Ana M; Figueroa, José I; Chavarrieta, Juan M; Viñuela, Elisa; Rebollar-Alviter, Ángel; Miranda, Mario A; Valle, Javier; Pineda, Samuel

    2017-07-18

    Control of the Asian citrus psyllid Diaphorina citri Kuwayama, the most important pest of citrus worldwide, is based on the use of insecticides, though unsatisfactory results have recently been reported. In this study, insecticide resistance of D. citri to three insecticides (bifenthrin, malathion, and chlorpyrifos) was examined. Three populations (designated Dci-CParácuaro, Dci-El Junco, and Dci-Antúnez) of both adults and fourth-instar D. citri individuals were collected in 2014 at two different times and on one occasion, respectively, from three locations (Crucero de Parácuaro, El Junco, and Antúnez). These locations represent the major commercial Mexican lemon production areas in the Apatzingán Valley in the state of Michoacán, Mexico. The three populations of D. citri adults and fourth-instar nymphs at the different collection times showed low levels of resistance (≤7-fold) to bifenthrin, but were very resistant to malathion (≤345- and ≤432-fold for adults and fourth instars, respectively) and chlorpyrifos (≤2435- and ≤1424-fold for adults and fourth instars, respectively). Resistance levels to the tested insecticides were highly variable but homogeneous among seasons and localities. Resistance management programmes that include crop sanitation, use of biological and cultural control practices, and rotation of insecticide classes should be established, particularly in areas where D. citri has developed resistance to malathion and chlorpyrifos. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  14. Metabolic Interplay between the Asian Citrus Psyllid and Its Profftella Symbiont: An Achilles' Heel of the Citrus Greening Insect Vector.

    Directory of Open Access Journals (Sweden)

    John S Ramsey

    Full Text Available 'Candidatus Liberibacter asiaticus' (CLas, the bacterial pathogen associated with citrus greening disease, is transmitted by Diaphorina citri, the Asian citrus psyllid. Interactions among D. citri and its microbial endosymbionts, including 'Candidatus Profftella armatura', are likely to impact transmission of CLas. We used quantitative mass spectrometry to compare the proteomes of CLas(+ and CLas(- populations of D. citri, and found that proteins involved in polyketide biosynthesis by the endosymbiont Profftella were up-regulated in CLas(+ insects. Mass spectrometry analysis of the Profftella polyketide diaphorin in D. citri metabolite extracts revealed the presence of a novel diaphorin-related polyketide and the ratio of these two polyketides was changed in CLas(+ insects. Insect proteins differentially expressed between CLas(+ and CLas(- D. citri included defense and immunity proteins, proteins involved in energy storage and utilization, and proteins involved in endocytosis, cellular adhesion, and cytoskeletal remodeling which are associated with microbial invasion of host cells. Insight into the metabolic interdependence between the insect vector, its endosymbionts, and the citrus greening pathogen reveals novel opportunities for control of this disease, which is currently having a devastating impact on citrus production worldwide.

  15. Laser system for identification, tracking, and control of flying insects.

    Science.gov (United States)

    Mullen, Emma R; Rutschman, Phillip; Pegram, Nathan; Patt, Joseph M; Adamczyk, John J; Johanson, 3ric

    2016-05-30

    Flying insects are common vectors for transmission of pathogens and inflict significant harm to humans and agricultural production in many parts of the world. We present proof of principle for an optical system capable of highly specific vector control. This system utilizes a combination of optical sources, detectors, and sophisticated software to search, detect, and identify flying insects in real-time, with the capability of eradication using a lethal laser pulse. We present data on two insect species to show species distinction; Diaphorina citri, a vector of the causal agent of citrus greening disease, and Anopheles stephensi, a malaria vector.

  16. Compuestos volátiles de brotes de limón persa y limón mexicano asociados a síntomas del HLB (Huanglongbing).

    OpenAIRE

    Mendoza Peña, Estrella

    2014-01-01

    El Huanglongbing (HLB) es una enfermedad de los cítricos que se caracteriza por causar áreas moteadas en hojas, generar frutos deformes y decolorados, semillas abortivas, causar pérdida paulatina de la producción y provocar que la muerte de los árboles infectados sea inevitable. El agente asociado a esta enfermedad es la bacteria Candidatus Liberibacter ssp., y en México es dispersada por el vector Diaphorina citri Kuwayama (Hemiptera: Liviidae). En el presente estudio, mediante el método de ...

  17. HPLC Determination of Naringin, Hesperidin and Neohesperidin in Citri Reticulatae Pericarpium and Aurantii Fructus Extract%HPLC测定陈皮-枳壳药对提取物中柚皮苷、橙皮苷和新橙皮苷的含量

    Institute of Scientific and Technical Information of China (English)

    郭琦丽; 吕武清; 曾莉萍; 李才堂

    2012-01-01

    目的:建立陈皮-枳壳药对提取物中柚皮苷、橙皮苷和新橙皮苷的含量测定方法.方法:采用HPLC,Agilent Hypersill C18(4.0 mm×250 mm,5μm)色谱柱,以乙腈-0.1%磷酸(24∶ 76)为流动相,流速1 mL· min-1,检测波长283 nm,柱温40℃.结果:柚皮苷、橙皮苷和新橙皮分别在6.05 ~60.5,4.34 ~ 43.4,7.14 ~71.40 mg·L-1线性关系良好(r均>0.999 5);加样回收率分别为98.65%,98.66%,100.78%,其RSD分别为2.01%,1.52%,2.11%;重复性RSD分别为2.41%,1.56%,2.52%(n=6).结论:该方法简便可行,重复性好,结果准确可靠,可用于陈皮-枳壳药对提取物中柚皮苷、橙皮苷和新橙皮苷的含量测定.%Objective: To develop a method to determine naringin, hesperidin and neohesperidin in Citri Reticulatae Pericarpium and Aurantii Fructus. Method; HPLC was used to determine directly naringin, hesperidin and neohesperidin of Citri Reticulatae Pericarpium and Aurantii Fructus. Phenomsil C18 (4. 0 mm x 250 mm, 5 μm) as the chromatographic column, and acetonitrile-10% aqueous phosphoric acid (24:76) as the mobile phase, the flow rate of 1.0 mL -min , the column temperature was at 40 °C. Result; The linearity of verbascoside was good in the range of 6. 05-60. 5 , 4. 3443. 4 and 7. 14-71. 40 mg - L-1 ( r > 0. 999 5 ) . The recovery of the added sample was 98. 65% , 98. 66% and 100. 78% with RSD of 2. 01% , 1. 52% and 2. 11% . Conclusion.- The method is simple, accurate and can be used for the determination of naringin, hesperidin and neohesperidin in Tangerine peel and Aurantii Fructus extract.

  18. Oral delivery of double-stranded RNAs and siRNAs induces RNAi effects in the potato/tomato psyllid, Bactericerca cockerelli.

    Directory of Open Access Journals (Sweden)

    Hada Wuriyanghan

    Full Text Available The potato/tomato psyllid, Bactericerca cockerelli (B. cockerelli, and the Asian citrus psyllid, Diaphorina citri (D. citri, are very important plant pests, but they are also vectors of phloem-limited bacteria that are associated with two devastating plant diseases. B. cockerelli is the vector of Candidatus Liberibacter psyllaurous (solanacearum, which is associated with zebra chip disease of potatoes, and D. citri is the vector of Ca. Liberibacter asiaticus, which is associated with the Huanglongbing (citrus greening disease that currently threatens the entire Florida citrus industry. Here we used EST sequence information from D. citri to identify potential targets for RNA interference in B. cockerelli. We targeted ubiquitously expressed and gut-abundant mRNAs via injection and oral acquisition of double-stranded RNAs and siRNAs and were able to induce mortality in recipient psyllids. We also showed knockdown of target mRNAs, and that oral acquisition resulted primarily in mRNA knockdown in the psyllid gut. Concurrent with gene knockdown was the accumulation of target specific ∼ 21 nucleotide siRNAs for an abundant mRNA for BC-Actin. These results showed that RNAi can be a powerful tool for gene function studies in psyllids, and give support for continued efforts for investigating RNAi approaches as possible tools for psyllid and plant disease control.

  19. Sclerenchymatous ring as a barrier to phloem feeding by Asian citrus psyllid: Evidence from electrical penetration graph and visualization of stylet pathways.

    Science.gov (United States)

    George, Justin; Ammar, El-Desouky; Hall, David G; Lapointe, Stephen L

    2017-01-01

    Asian citrus psyllid (Diaphorina citri) feeding behaviors play a significant role in the transmission of the phloem-limited Candidatus Liberibacter asiaticus (CLas) bacterium that causes the economically devastating citrus greening disease. Sustained phloem ingestion by D. citri on CLas infected plants is required for pathogen acquisition and transmission. Recent studies have shown a fibrous ring of thick-walled sclerenchyma around the phloem in mature, fully expanded citrus leaves that is more prominent on the abaxial compared with the adaxial side. The composition and thickness of this fibrous ring may have an important role in selection of feeding sites by D. citri based on leaf age and leaf surface, which in turn can affect pathogen acquisition and transmission. We measured feeding behavior using electrical penetration graph (EPG) recordings of individual D. citri adults placed on abaxial or adaxial surfaces of young or mature Valencia orange leaves to study the role of the sclerenchymatous ring in modifying D. citri feeding behavior. Feeding sites on the same leaf tissues were then sectioned and examined by epifluorescence microscopy. The duration of phloem ingestion (E2 waveform) by psyllids was significantly reduced on mature compared with young leaves, and on abaxial compared with adaxial leaf surfaces. The longest duration of phloem ingestion was observed from psyllids placed on the adaxial side of young leaves that had the least developed sclerenchyma. Bouts of phloem salivation (E1 waveform), however, were significantly longer on mature leaves compared with young leaves. D. citri adults made consecutive phloem feeding attempts (bouts) on the abaxial side of mature leaves and those bouts resulted in unsuccessful or shorter periods of phloem ingestion. Adults also made more frequent and longer bouts of xylem ingestion on mature leaves compared with adult psyllids placed on young leaves. Epifluorescence microscopy showed that the fibrous ring in young

  20. Induced release of a plant-defense volatile 'deceptively' attracts insect vectors to plants infected with a bacterial pathogen.

    Directory of Open Access Journals (Sweden)

    Rajinder S Mann

    Full Text Available Transmission of plant pathogens by insect vectors is a complex biological process involving interactions between the plant, insect, and pathogen. Pathogen-induced plant responses can include changes in volatile and nonvolatile secondary metabolites as well as major plant nutrients. Experiments were conducted to understand how a plant pathogenic bacterium, Candidatus Liberibacter asiaticus (Las, affects host preference behavior of its psyllid (Diaphorina citri Kuwayama vector. D. citri were attracted to volatiles from pathogen-infected plants more than to those from non-infected counterparts. Las-infected plants were more attractive to D. citri adults than non-infected plants initially; however after feeding, psyllids subsequently dispersed to non-infected rather than infected plants as their preferred settling point. Experiments with Las-infected and non-infected plants under complete darkness yielded similar results to those recorded under light. The behavior of psyllids in response to infected versus non-infected plants was not influenced by whether or not they were carriers of the pathogen. Quantification of volatile release from non-infected and infected plants supported the hypothesis that odorants mediate psyllid preference. Significantly more methyl salicylate, yet less methyl anthranilate and D-limonene, was released by infected than non-infected plants. Methyl salicylate was attractive to psyllids, while methyl anthranilate did not affect their behavior. Feeding on citrus by D. citri adults also induced release of methyl salicylate, suggesting that it may be a cue revealing location of conspecifics on host plants. Infected plants were characterized by lower levels of nitrogen, phosphorus, sulfur, zinc, and iron, as well as, higher levels of potassium and boron than non-infected plants. Collectively, our results suggest that host selection behavior of D. citri may be modified by bacterial infection of plants, which alters release of

  1. Antifungal activity of chitosan(oligochitosan) on Geotrichum candidum and Alternaria citri of citrus and postharvest disease control%壳聚(寡)糖对柑橘酸腐、黑腐病菌的抑制作用及对采后病害的防治

    Institute of Scientific and Technical Information of China (English)

    王丽; 赵盼; 孟祥红

    2011-01-01

    In vitro,the pore germination,germ tube length and mycelial growth of the two fungi were used to evaluate to the effect of chitosan and oligochitosan. The effects of chitosan and ologochitosan on disease control were investigated by stab inoculation. The results showed that chitosan and oligochitosan at 10mg/mL almost completely inhibited the mycelial growth of the two fungi. Chitosan and oligochitosan at 0. 5mg/mL completely inhibited the spore germination and germ tube elongation of Geotrichum candidum. Moreover,2mg/mL of chitosan could completely inhibit the spore germination and germ tube elongation of Alternaria citri,but 2mg/mL of oligochitosan had a lower inhibitory effect. In vivo results showed that the control effect of chitosan and oligochitosan at 1% on fruit disease were better than lower concentration.%通过体外实验测定不同浓度的壳聚糖和壳寡糖对酸腐和黑腐病菌孢子萌发、芽管长度和菌丝生长的抑制作用;采用刺伤接种方法评价壳聚糖和壳寡糖对柑橘果实两种病害的防治作用。离体实验结果表明,壳聚糖和壳寡糖浓度为10mg/mL时几乎完全抑制两种病原菌的菌丝生长;浓度为0.5mg/mL壳聚糖和壳寡糖能完全抑制柑橘酸腐病菌(Geotrichum candidum)孢子萌发和芽管伸长的效果,而2mg/mL的壳聚糖才能完全抑制柑橘黑腐病菌(Alternaria citri)孢子萌发和芽管伸长,但相同浓度的壳寡糖抑制A.citri的孢子萌发和芽管伸长效果较差。体内实验结果表明,其病害防治效果具有浓度依赖性,高浓度(1%)的壳聚糖和壳寡糖对果实病害的防治效果较低浓度的防治效果好。

  2. Repellency of selected biorational insecticides to potato psyllid, Bactericera cockerelli (Hemiptera: Psyllidae)

    Science.gov (United States)

    Bactericera cockerelli has recently become a major concern because of its direct feeding and vectoring of bacterial diseases in many solanaceous crops. The repellency of four biorational insecticides, MOI-201 (a Chinese medicine plant extract), Requiem (a plant extract of Chenopodium ambrosioides), ...

  3. Ecological and genetic differences between Cacopsylla melanoneura (Hemiptera, Psyllidae populations reveal species host plant preference.

    Directory of Open Access Journals (Sweden)

    Valeria Malagnini

    Full Text Available The psyllid Cacopsylla melanoneura is considered one of the vectors of 'Candidatus Phytoplasma mali', the causal agent of apple proliferation disease. In Northern Italy, overwintered C. melanoneura adults reach apple and hawthorn around the end of January. Nymph development takes place between March and the end of April. The new generation adults migrate onto conifers around mid-June and come back to the host plant species after overwintering. In this study we investigated behavioural differences, genetic differentiation and gene flow between samples of C. melanoneura collected from the two different host plants. Further analyses were performed on some samples collected from conifers. To assess the ecological differences, host-switching experiments were conducted on C. melanoneura samples collected from apple and hawthorn. Furthermore, the genetic structure of the samples was studied by genotyping microsatellite markers. The examined C. melanoneura samples performed better on their native host plant species. This was verified in terms of oviposition and development of the offspring. Data resulting from microsatellite analysis indicated a low, but statistically significant difference between collected-from-apple and hawthorn samples. In conclusion, both ecological and genetic results indicate a differentiation between C. melanoneura samples associated with the two host plants.

  4. A revision of the Chinese species of Cyamophiliopsis (Hemiptera: Psylloidea: Psyllidae) associated with Spiraea (Rosaceae).

    Science.gov (United States)

    Luo, Xinyu; Li, Fasheng; Cai, Wanzhi

    2015-03-20

    Cyamophiliopsis is a small genus restricted to the Palaearctic Region and associated with Spiraea spp. (Rosaceae). In the present work, following five species are recognized in China: C. pseudofasciata sp. nov., C. sarmatica, C. spinosa sp. nov., C. xinjiangana sp. nov. and C. zaisani. Cyamophiliopsis is diagnosed, redescribed in detail, and its phylogenetic relationships are discussed. All the species are described or redescribed, and the fifth instar immature of C. zaisani is described for the first time. Nomenclatorial problems are discussed concerning the Far East Russian Psylla spiraee which is transferred to Cyamophiliopsis as C. spiraee comb. nov.

  5. Insecticidal activity of neem oil against Gyropsylla spegazziniana (Hemiptera: Psyllidae) nymphs on Paraguay tea seedlings.

    Science.gov (United States)

    Formentini, M A; Alves, L F A; Schapovaloff, M E

    2016-01-01

    Gyropsylla spegazziniana (Paraguay tea ampul) is one of the most important pests of Paraguay tea plants, and prohibition of synthetic insecticide use for control of this pest has led to the search for alternative methods. This laboratory study aimed to compare different control strategies for G. spegazziniana, utilizing a commercial neem seed oil product. Paraguay tea seedlings were treated with neem oil solution both pre- and post-infestation with 5th instar nymphs. The systemic action of neem oil was also evaluated by treating plant soil with the neem oil solution, followed by transfer of the insects to plants 24 h post-treatment. Spray treatments were effective against the pest, especially post-infestation (80% mortality), demonstrating the potential of neem oil for control of the Paraguay tea ampul. No significant effects were observed with respect to systemic activity.

  6. First records of Acizzia jamatonica (Kuwayama) and Glycaspis brimblecombei Moore, (Hemiptera: Psyllidae, Aphalaridae) in Montenegro

    OpenAIRE

    Malumphy, Chris; Perović, Tatjana; Hrnčić, Snježana; RADONJIĆ, SANJA; RAIČEVIĆ, MILORAD

    2013-01-01

    Albizia psyllid, Acizzia jamatonica (Kuwayama), and red gum lerp psyllid, Glycaspis brimblecombei Moore, were found in Montenegro for the first time during 2009 and 2012 respectively. Acizzia jamatonica is native to East Asia, and has spread rapidly in Europe since it was first reported from Italy in 2002. It was first collected from Albizzia julibrissin in Podgorica, September 2009, and subsequently from Albizzia sp. and A. julibrissin in Herceg Novi, October 2010, and June and October 2012,...

  7. Insecticidal activity of neem oil against Gyropsylla spegazziniana (Hemiptera: Psyllidae nymphs on Paraguay tea seedlings

    Directory of Open Access Journals (Sweden)

    M. A. Formentini

    Full Text Available Abstract Gyropsylla spegazziniana (Paraguay tea ampul is one of the most important pests of Paraguay tea plants, and prohibition of synthetic insecticide use for control of this pest has led to the search for alternative methods. This laboratory study aimed to compare different control strategies for G. spegazziniana, utilizing a commercial neem seed oil product. Paraguay tea seedlings were treated with neem oil solution both pre- and post-infestation with 5th instar nymphs. The systemic action of neem oil was also evaluated by treating plant soil with the neem oil solution, followed by transfer of the insects to plants 24 h post-treatment. Spray treatments were effective against the pest, especially post-infestation (80% mortality, demonstrating the potential of neem oil for control of the Paraguay tea ampul. No significant effects were observed with respect to systemic activity.

  8. Development of RLK-Derived Molecular Markers Associating with the Resistance to Citrus Canker [Xanthomonas axonopodis pv. Citri (Xac)] Disease%开发与柑桔抗溃疡病基因连锁的由RLK-RGC衍生的分子标记

    Institute of Scientific and Technical Information of China (English)

    向旭; 郑启发; 黄舒; 陈存贤; Frederick G. Gmitter Jr.; 邓占鳌

    2005-01-01

    本文是在先期研究工作的基础上,利用从柑桔与枳属间杂种的基因组DNA所获得的类受体激酶(RLK)的候选抗病基因序列,重新设计引物,对柑桔抗溃疡病材料和感病材料开展以PCR扩增为基础的对比分析,其中一对引物‘19h16/DdeI'的扩增产物经限制性内切酶DdeI酶切,揭示了抗性材料(Ichang Papeda,Meiwa kumquat,Maruhi kumquat and Nagami kumquat)和感病材料(Flying Dragon,Valencia orange and Palestine sweetlime)之间的多态性差异,同时,在C.Ichangensis的自交F1代和Palestine sweetlime× Ichang Papeda的杂交F1代群体中也见明显的多态性差异;对这些F1代个体进行溃疡病病原[Xanthomonas axonopodis pv.Citri(Xac)]接种作抗病性表型鉴定,结果表明,该分子标记‘19h16/DdeI'与柑桔溃疡病抗性密切相关;经纯化测序的结果进一步证明,抗性材料PCR扩增产物具有完全一致的序列和DdeI酶切位点,但感病材料缺少这一DdeI酶切位点;染色体步行(primerwalking)在BAC文库的对应单克隆中获得了一个完整的抗病基因序列,与以前获得的2个抗病基因序列‘17o6RLKP'和‘26m19RLKP'相比,‘19h16RLKP'也具有Xa21抗病基因蛋白的所有特征,包括含有一个信号肽、同样数目的亮氨酸重复序列、跨膜域和激酶域等.基于该序列的开放读码框,发展了特异性更强可靠性更高的另一分子标记,在今后的研究工作中具有较大的应用潜力.%By utilizing the receptor-like kinase (RLK) RGC sequences previously obtained from the genomic DNA of an intergeneric Citrus and Poncirus hybrid, PCR amplification with one pair of new-designed primers was found to reveal polymorphism between the theoretical resistant species (Ichang Papeda, Meiwa kumquat,Maruhi kumquat and Nagami kumquat) and susceptible species (Flying Dragon, Valencia orange and Palestine sweet lime ) and among the self-crossing population of C. Ichangensis and F1 population of Palestine sweet lime

  9. Plant pathogen-induced volatiles attract parasitoids to increase parasitism of an insect vector

    Directory of Open Access Journals (Sweden)

    Xavier eMartini

    2014-05-01

    Full Text Available Interactions between plant pathogens and arthropods have been predominantly studied through the prism of herbivorous arthropods. Currently, little is known about the effect of plant pathogens on the third trophic level. This question is particularly interesting in cases where pathogens manipulate host phenotype to increase vector attraction and presumably increase their own proliferation. Indeed, a predator or a parasitoid of a vector may take advantage of this manipulated phenotype to increase its foraging performance. We explored the case of a bacterial pathogen, Candidatus Liberibacter asiaticus (Las, which modifies the odors released by its host plant (citrus trees to attract its vector, the psyllid Diaphorina citri. We found that the specialist parasitoid of D. citri, Tamarixia radiata, was attracted more toward Las-infected than uninfected plants. We demonstrated that this attractiveness was due to the release of methyl salicylate. Parasitization of D. citri nymphs on Las-infected plants was higher than on uninfected controls. Also, parasitization was higher on uninfected plants baited with methyl salicylate than on non-baited controls. This is the first report of a parasitoid ‘eavesdropping’ on a plant volatile induced by bacterial pathogen infection, which also increases effectiveness of host seeking behavior of its herbivorous vector.

  10. Effect of Cyantraniliprole, a Novel Insecticide, on the Inoculation of Candidatus Liberibacter Asiaticus Associated with Citrus Huanglongbing by the Asian Citrus Psyllid (Hemiptera: Liviidae).

    Science.gov (United States)

    Ammar, El-Desouky; Hall, David G; Alvarez, Juan M

    2015-04-01

    The Asian citrus psyllid (Diaphorina citri Kuwayama) is the principal vector of 'Candidatus Liberibacter asiaticus' (CLas) associated with huanglongbing (HLB), the most serious citrus disease worldwide. New control measures including pesticides are urgently needed to combat HLB, especially to protect young or newly planted citrus trees from CLas-inoculation by vector psyllids. Here, we tested CLas-inoculation by D. citri adults (CLas-exposed, reared on infected plants) by feeding them for 7 d on excised healthy citrus leaves with dry residues of cyantraniliprole (Exirel), a novel insecticide, in comparison with fenpropathrin (Danitol 2.4EC), an insecticide commonly used against D. citri. Fewer adults settled (putatively feeding or probing) on leaves treated with cyantraniliprole than those treated with fenpropathrin or water controls. Also, psyllid adults died at a slower rate on leaves treated with cyantraniliprole than those treated with fenpropathrin, although the final cumulative mortality did not differ between the two treatments. In quantitative real-time polymerase chain reaction tests, 59.0-65.3% of the CLas-exposed psyllid adults were proven to be CLas-positive. Inoculation rates of CLas (using 10 adults per leaf) into untreated healthy citrus leaves (47.5-85%) were significantly higher than rates into leaves treated with cyantraniliprole or fenpropathrin (2.5-12.5%). Reduced inoculation rates to leaves treated with cyantraniliprole probably occurred as a result of reduced feeding or probing by D. citri. The excised leaf assay method, which took only a few weeks compared with up to a year or longer using whole plants, can be an effective tool for testing the effect of new pesticides or other treatments in reducing CLas inoculation or transmission by psyllid vectors. Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by a US Government employees and is in the public domain in the US.

  11. PENDIDIKAN DAN PELATIHAN PENGENDALIAN TERPADU PENYAKIT CVPD (CITRUS VEIN PHLOEM DEGENERATION PADA PERTANAMAN JERUK DI DESA PELAGA, KECAMATAN PETANG, KABUPATEN BADUNG

    Directory of Open Access Journals (Sweden)

    WAYAN ADIARTAYASA

    2012-09-01

    Full Text Available ABSTRACT Education and training on integrated pest management (IPM of CVPD (Citrus Vein Phloem Degeneration was held at Pelaga village, Badung regency on 22 September 2005. The activities aimed to improve knowledge of the farmers about CVPD and factors influencing the intensity, insect vector and integrated pest management of CVPD. The topics convered were disease symptons, and pathogen bacteria, Diaphorina citri are as vector insect infection mechanism, disease distribubution and integrated management of CVPD. The training was attended by 26 participants from local groups of Pelaga village. The method used in activities were lectures, demonstration and practical work in the citrus field. All participants enthusiastically took part and hoped to have the next intensive traning of citrus culture.

  12. PENDIDIKAN DAN PELATIHAN PEMBIBITAN JERUK BEBAS PENYAKIT CVPD (Citrus Vein Phloem Degeneration DI DESA KATUNG KINTAMANI

    Directory of Open Access Journals (Sweden)

    Adiartayasa W

    2012-10-01

    Full Text Available The community service in the form of training about free orange seed of disease CVPD (Citrus Vein Phloem Degenaration was conducted in Katung village, district of Kintamani, Bangli regency on 10 August 2007. The training activity aimed to improve knowledge of farmers about CVPD disease, symptom, transmission, insect vector, and integrated pest management. The topics covered were disease symptoms, and pathogen bacteria, Diaphorina citri are as insect vector infection mechanism, election of free orange mains crops CVPD, disease distribution and integrated management of CVPD. The training was attended by 25 participants from local groups of Katung village. The methods used in this activity were lectures, demonstration and practical work in the citrus field. All participants enthusiastically took part and hoped to have the next intensive training of citrus culture.

  13. Candidate gene markers for Candidatus Liberibacter asiaticus for detecting citrus greening disease.

    Science.gov (United States)

    Nageswara-Rao, Madhugiri; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame

    2013-06-01

    Citrus Huanglongbing (HLB) also known as citrus greening is one of the most devastating diseases of citrus worldwide. The disease is caused by Candidatus Liberibacter bacterium, vectored by the psyllid Diaphorina citri Kuwayama and Trioza erytreae Del Guercio. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection. The aim of this study was to develop effective gene-specific primer pairs for polymerase chain reaction based method for quick screening of HLB disease. Thirty-two different gene-specific primer pairs, across the Ca. Liberibacter asiaticus genome, were successfully developed. The possibility of these primer pairs for cross-genome amplification across 'Ca. Liberibacter africanus' and 'Ca. Liberibacter americanus' were tested. The applicability of these primer pairs for detection and differentiation of Ca Liberibacter spp. is discussed.

  14. 药剂防控柑橘黄龙病的探讨%Discussion on Pesticides Controlling the Citrus Huanglongbing

    Institute of Scientific and Technical Information of China (English)

    成家壮

    2011-01-01

    Based on the characteristics and the epidemic law of citrus disease,an idea was put forwardresearching and developing the antifeedant against diaphorina citri and the bactericides to control pathogenic bacteria andto bukme the host plant resistance in order to control Huanglongbing(HLB),at the same time,the relevant issues were alsodiscussed.%根据柑橘黄龙病的发生特点与流行规律,提出了研究开发柑橘木虱拒食剂和抑制病原菌以及可诱导寄主抗病性的杀菌剂用以防控柑橘黄龙病的构想,并对相关问题进行了探讨.

  15. Odor coding in a disease-transmitting herbivorous insect, the Asian citrus psyllid.

    Science.gov (United States)

    Coutinho-Abreu, Iliano V; McInally, Shane; Forster, Lisa; Luck, Robert; Ray, Anandasankar

    2014-07-01

    Olfactory systems discriminate odorants very efficiently and herbivorous insects use them to find hosts in confounding and complex odor landscapes. The Asian citrus psyllid (ACP), Diaphorina citri, feeds on citrus flush and transmits Candidatus Liberibacter that causes citrus greening disease globally. Here, we perform a systematic analysis of odor detection in the ACP antenna using single-unit electrophysiology of rhinarial plate sensilla to a large panel of odorants from plants. We identify neurons that respond strongly to odorants found in the host citrus plants. Comparisons with the generalist yeast-feeding Drosophila melanogaster and specialist anthropophilic Anopheles gambiae reveal differences in odor-coding strategies for the citrus-seeking ACP. These findings provide a foundation for understanding host-odor coding in herbivorous insects.

  16. Monitoring and controlling measures of Huanglongbin disease in Guanxi pomelo plant%琯溪蜜柚黄龙病的监测与防控

    Institute of Scientific and Technical Information of China (English)

    杨文火

    2012-01-01

    通过对采集的琯溪蜜柚的各种典型黄花症状样品进行黄龙病病原PCR测定,在分析病害症状并进行实时监测的基础上,提出及时铲除病株和防治木虱、严格控制外来种苗的进入等防控措施。%Pathogen of Citrus Huanglongbing(HLB)was detected by PCR techque by using the samples of Guanxi pomelo,which showed typical leaf-yellowing symptom.Based on analyzing the symptom of HLB and on-the-spot monitoring to the disease,we put forward a set of integrated control techniques,including removing HLB-affected plants,controlling Diaphorina citri,strictly forbiding the entrance of alien seedings.

  17. First records of parasitoids attacking the Asian citrus psyllid in Ecuador

    Directory of Open Access Journals (Sweden)

    Diego E. Portalanza

    Full Text Available ABSTRACT First records of parasitoids attacking the Asian citrus psyllid in Ecuador. The objective of the current study was to investigate the presence of natural enemies of Diaphorina citri (Hemiptera: Liviidae (the Asian citrus psyllid in Ecuador. Incidence of parasitoid Diaphorencyrtus aligarhensis (Hymenoptera: Encyrtidae was assessed between November 2015 and March 2016, in Letamendi, Febres-Cordero and Tarqui, urban districts of Guayaquil. Highest incidence of parasitism occurred in those regions and seasons of the year with the highest temperatures commensurate with increase of citrus plant shoots. Similar to their host, these parasitoids appear to have established in Ecuador by accident, and were not the result of purposeful introduction. This fortuitous introduction is a potentially helpful tool in controlling the Asian citrus psyllid, and potentially Huanglongbing.

  18. Modelling the effect of an alternative host population on the spread of citrus Huanglongbing

    Science.gov (United States)

    d'A. Vilamiu, Raphael G.; Ternes, Sonia; Laranjeira, Francisco F.; de C. Santos, Tâmara T.

    2013-10-01

    The objective of this work was to model the spread of citrus Huanglongbing (HLB) considering the presence of a population of alternative hosts (Murraya paniculata). We developed a compartmental deterministic mathematical model for representing the dynamics of HLB disease in a citrus orchard, including delays in the latency and incubation phases of the disease in the plants and a delay period on the nymphal stage of Diaphorina citri, the insect vector of HLB in Brazil. The results of numerical simulations indicate that alternative hosts should not play a crucial role on HLB dynamics considering a typical scenario for the Recôncavo Baiano region in Brazil . Also, the current policy of removing symptomatic plants every three months should not be expected to significantly hinder HLB spread.

  19. Candidate gene markers for Candidatus Liberibacter asiaticus for detecting citrus greening disease

    Indian Academy of Sciences (India)

    Madhugiri Nageswara-Rao; Mike Irey; Stephen M Garnsey; Siddarame Gowda

    2013-06-01

    Citrus Huanglongbing (HLB) also known as citrus greening is one of the most devastating diseases of citrus worldwide. The disease is caused by Candidatus Liberibacter bacterium, vectored by the psyllid Diaphorina citri Kuwayama and Trioza erytreae Del Guercio. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection. The aim of this study was to develop effective gene-specific primer pairs for polymerase chain reaction based method for quick screening of HLB disease. Thirty-two different gene-specific primer pairs, across the Ca. Liberibacter asiaticus genome, were successfully developed. The possibility of these primer pairs for cross-genome amplification across `Ca. Liberibacter africanus’ and `Ca. Liberibacter americanus’ were tested. The applicability of these primer pairs for detection and differentiation of Ca Liberibacter spp. is discussed.

  20. Acquisition, Replication and Inoculation of Candidatus Liberibacter asiaticus following Various Acquisition Periods on Huanglongbing-Infected Citrus by Nymphs and Adults of the Asian Citrus Psyllid.

    Science.gov (United States)

    Ammar, El-Desouky; Ramos, John E; Hall, David G; Dawson, William O; Shatters, Robert G

    2016-01-01

    The Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), is the primary vector of Candidatus Liberibacter asiaticus (Las) implicated as causative agent of citrus huanglongbing (citrus greening), currently the most serious citrus disease worldwide. Las is transmitted by D. citri in a persistent-circulative manner, but the question of replication of this bacterium in its psyllid vector has not been resolved. Thus, we studied the effects of the acquisition access period (AAP) by nymphs and adults of D. citri on Las acquisition, multiplication and inoculation/transmission. D. citri nymphs or adults (previously non-exposed to Las) were caged on Las-infected citrus plants for an AAP of 1, 7 or 14 days. These 'Las-exposed' psyllids were then transferred weekly to healthy citrus or orange jasmine plants, and sampled via quantitative polymerase chain reaction (qPCR) analysis 1-42 days post-first access to diseased plants (padp); all tested nymphs became adults 7-14 days padp. Our results indicate that following 1 or 7 day AAP as nymphs 49-59% of Las-exposed psyllids became Las-infected (qPCR-positive), whereas only 8-29% of the psyllids were infected following 1-14 day AAP as adults. Q-PCR analysis also indicated that Las titer in the Las-exposed psyllids (relative to that of the psyllid S20 ribosomal protein gene) was: 1) significantly higher, and increasing at a faster rate, following Las acquisition as nymphs compared to that following Las acquisition as adults; 2) higher as post-acquisition time of psyllids on healthy plants increased reaching a peak at 14-28 days padp for nymphs and 21-35 days padp for adults, with Las titer decreasing or fluctuating after that; 3) higher with longer AAP on infected plants, especially with acquisition as adults. Our results strongly suggest that Las multiplies in both nymphs and adults of D. citri but attains much higher levels in a shorter period of time post-acquisition when acquired by nymphs than when acquired by adults

  1. Acquisition, Replication and Inoculation of Candidatus Liberibacter asiaticus following Various Acquisition Periods on Huanglongbing-Infected Citrus by Nymphs and Adults of the Asian Citrus Psyllid.

    Directory of Open Access Journals (Sweden)

    El-Desouky Ammar

    Full Text Available The Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae, is the primary vector of Candidatus Liberibacter asiaticus (Las implicated as causative agent of citrus huanglongbing (citrus greening, currently the most serious citrus disease worldwide. Las is transmitted by D. citri in a persistent-circulative manner, but the question of replication of this bacterium in its psyllid vector has not been resolved. Thus, we studied the effects of the acquisition access period (AAP by nymphs and adults of D. citri on Las acquisition, multiplication and inoculation/transmission. D. citri nymphs or adults (previously non-exposed to Las were caged on Las-infected citrus plants for an AAP of 1, 7 or 14 days. These 'Las-exposed' psyllids were then transferred weekly to healthy citrus or orange jasmine plants, and sampled via quantitative polymerase chain reaction (qPCR analysis 1-42 days post-first access to diseased plants (padp; all tested nymphs became adults 7-14 days padp. Our results indicate that following 1 or 7 day AAP as nymphs 49-59% of Las-exposed psyllids became Las-infected (qPCR-positive, whereas only 8-29% of the psyllids were infected following 1-14 day AAP as adults. Q-PCR analysis also indicated that Las titer in the Las-exposed psyllids (relative to that of the psyllid S20 ribosomal protein gene was: 1 significantly higher, and increasing at a faster rate, following Las acquisition as nymphs compared to that following Las acquisition as adults; 2 higher as post-acquisition time of psyllids on healthy plants increased reaching a peak at 14-28 days padp for nymphs and 21-35 days padp for adults, with Las titer decreasing or fluctuating after that; 3 higher with longer AAP on infected plants, especially with acquisition as adults. Our results strongly suggest that Las multiplies in both nymphs and adults of D. citri but attains much higher levels in a shorter period of time post-acquisition when acquired by nymphs than when acquired by

  2. Primeiro registro de Glycaspis brimblecombei Moore 1964, (Hemiptera: Psyllidae em Eucalipto no Estado de Santa Catarina, Brasil First record of Glycaspis brimblecombei Moore 1964, (Hemiptera: Psyllidae in Eucalyptus in Santa Catarina State, Brazil

    Directory of Open Access Journals (Sweden)

    Junir Antonio Lutinski

    2006-04-01

    Full Text Available Conhecido como psilídeo-de-concha, Glycaspis brimblecombei é um inseto sugador de origem australiana e ataca várias espécies de eucalipto. Sua presença no Brasil foi registrada pela primeira vez no Estado de São Paulo em junho de 2003. Posteriormente, sua ocorrência foi registrada também para os Estados de Minas Gerais, Goiás, Paraná e Mato Grosso do Sul. As ninfas possuem coloração amarelada e encontram-se protegidas sob pequenas conchas enquanto os adultos, com coloração marrom-alaranjado a amarelo-esverdeado, medem cerca de três milímetros de comprimento. Dentre os principais danos que causam às plantas estão a queda prematura das folhas, redução da área fotossintética e superbrotamento. Em alguns casos, pode levar à morte da planta. No Estado de Santa Catarina, o G. brimblecombei foi encontrado atacando algumas plantas de Eucalyptus tereticornis localizadas no pátio de uma empresa no perímetro urbano da cidade de Chapecó.Known as Red gum lerp psyllid, Glycaspis brimblecombei is a sucker insect of Australian origin whose attacks several eucalyptus species. Its presence in Brazil was reported for the first time in the São Paulo State in June, 2003. Later, its occurrence was also registered in Minas Gerais, Goiás, Paraná and Mato Grosso do Sul States. The nymphs possess yellowish color and they are protected under small shells while the adults have brown-orange to yellow-greenish color, and about three milimeters. Among the main damages that causes to the plants are the premature leaves fall and reduction of the photosynthetic area. In some cases it can occurs plant death. In Santa Catarina State it was found attacking some trees of Eucalyptus tereticornis in the courtyard of a company in the urban perimeter of the Chapecó town.

  3. Invasive mutualisms between a plant pathogen and insect vectors in the Middle East and Brazil.

    Science.gov (United States)

    Queiroz, Renan Batista; Donkersley, Philip; Silva, Fábio Nascimento; Al-Mahmmoli, Issa Hashil; Al-Sadi, Abdullah Mohammed; Carvalho, Claudine Márcia; Elliot, Simon L

    2016-12-01

    Complex multi-trophic interactions in vectorborne diseases limit our understanding and ability to predict outbreaks. Arthropod-vectored pathogens are especially problematic, with the potential for novel interspecific interactions during invasions. Variations and novelties in plant-arthropod-pathogen triumvirates present significant threats to global food security. We examined aspects of a phytoplasma pathogen of citrus across two continents. 'Candidatus Phytoplasma aurantifolia' causes Witches' Broom Disease of Lime (WBDL) and has devastated citrus production in the Middle East. A variant of this phytoplasma currently displays asymptomatic or 'silent' infections in Brazil. We first studied vector capacity and fitness impacts of the pathogen on its vectors. The potential for co-occurring weed species to act as pathogen reservoirs was analysed and key transmission periods in the year were also studied. We demonstrate that two invasive hemipteran insects-Diaphorina citri and Hishimonus phycitis-can vector the phytoplasma. Feeding on phytoplasma-infected hosts greatly increased reproduction of its invasive vector D. citri both in Oman and Brazil; suggesting that increased fitness of invasive insect vectors thereby further increases the pathogen's capacity to spread. Based on our findings, this is a robust system for studying the effects of invasions on vectorborne diseases and highlights concerns about its spread to warmer, drier regions of Brazil.

  4. Citrus huanglongbing: validation of Real-Time PCR (qPCR for the detection of Candidatus Liberibacter asiaticus and Candidatus Liberibacter americanus in Colombia

    Directory of Open Access Journals (Sweden)

    Jorge Evelio Ángel

    2014-12-01

    Full Text Available Citrus huanglongbing (HLB is the most destructive citrus disease. Two of the three known HLB-associated Candidatus Liberibacter species were recently found to be present in the Americas. In this study, eggs, nymphs and adults of Diaphorina citri Kuwayama (Hemiptera: Liviidae and suspect citrus plant materials were collected in 25 municipalities in the departments of Cundinamarca, Santander, Valle del Cauca, Meta and Quindio (Colombia. The detection sensitivity, specificity and assay performance of the 16S rDNA-based real-time PCR (qPCR were validated for the field survey of the disease in Colombia. The validation confirmed the reliability and robustness of the real-time PCR method for the detection of HLB bacteria in host citrus plant tissues and the vector D. citri. The diagnosis was performed for Candidatus Liberibacter asiaticus (Ca. L. asiaticus and for Candidatus Liberibacter americanus (Ca. L. americanus on 168 citrus plant material samples and 239 insect samples. Neither Ca. L. asiaticus nor Ca. L. americanus were detected in the host plants or insects vector, confirming the absence of the disease in the citrus-producing areas of Colombia.

  5. Evolutionary conservation of candidate osmoregulation genes in plant phloem sap-feeding insects.

    Science.gov (United States)

    Jing, X; White, T A; Luan, J; Jiao, C; Fei, Z; Douglas, A E

    2016-06-01

    The high osmotic pressure generated by sugars in plant phloem sap is reduced in phloem-feeding aphids by sugar transformations and facilitated water flux in the gut. The genes mediating these osmoregulatory functions have been identified and validated empirically in the pea aphid Acyrthosiphon pisum: sucrase 1 (SUC1), a sucrase in glycoside hydrolase family 13 (GH13), and aquaporin 1 (AQP1), a member of the Drosophila integral protein (DRIP) family of aquaporins. Here, we describe molecular analysis of GH13 and AQP genes in phloem-feeding representatives of the four phloem-feeding groups: aphids (Myzus persicae), coccids (Planococcus citri), psyllids (Diaphorina citri, Bactericera cockerelli) and whiteflies (Bemisia tabaci MEAM1 and MED). A single candidate GH13-SUC gene and DRIP-AQP gene were identified in the genome/transcriptome of most insects tested by the criteria of sequence motif and gene expression in the gut. Exceptionally, the psyllid Ba. cockerelli transcriptome included a gut-expressed Pyrocoelia rufa integral protein (PRIP)-AQP, but has no DRIP-AQP transcripts, suggesting that PRIP-AQP is recruited for osmoregulatory function in this insect. This study indicates that phylogenetically related SUC and AQP genes may generally mediate osmoregulatory functions in these diverse phloem-feeding insects, and provides candidate genes for empirical validation and development as targets for osmotic disruption of pest species. © 2016 The Authors. Insect Molecular Biology published by John Wiley & Sons Ltd on behalf of The Royal Entomological Society.

  6. Horizontal gene acquisition of Liberibacter plant pathogens from a bacteriome-confined endosymbiont of their psyllid vector.

    Directory of Open Access Journals (Sweden)

    Atsushi Nakabachi

    Full Text Available he Asian citrus psyllid Diaphorina citri is a notorious agricultural pest that transmits the phloem-inhabiting alphaproteobacterial 'Candidatus Liberibacter asiaticus' and allied plant pathogens, which cause the devastating citrus disease called Huanglongbing or greening disease. D. citri harbors two distinct bacterial mutualists in the symbiotic organ called bacteriome: the betaproteobacterium 'Candidatus Profftella armatura' in the syncytial cytoplasm at the center of the bacteriome, and the gammaproteobacterium 'Candidatus Carsonella ruddii' in uninucleate bacteriocytes. Here we report that a putative amino acid transporter LysE of Profftella forms a highly supported clade with proteins of L. asiaticus, L. americanus, and L. solanacearum. L. crescens, the most basal Liberibacter lineage currently known, lacked the corresponding gene. The Profftella-Liberibacter subclade of LysE formed a clade with proteins from betaproteobacteria of the order Burkholderiales, to which Profftella belongs. This phylogenetic pattern favors the hypothesis that the Liberibacter lineage acquired the gene from the Profftella lineage via horizontal gene transfer (HGT after L. crescens diverged from other Liberibacter lineages. K A/K S analyses further supported the hypothesis that the genes encoded in the Liberibacter genomes are functional. These findings highlight the possible evolutionary importance of HGT between plant pathogens and their insect vector's symbionts that are confined in the symbiotic organ and seemingly sequestered from external microbial populations.

  7. Elicitors of Host Plant Defenses Partially Suppress Pear Psylla (Cacopsylla pyricola, Hemiptera: Psyllidae) Populations under Field Conditions

    Science.gov (United States)

    Defense elicitors are products that activate acquired defense responses in plants, thus rendering the plants less susceptible to attack by a broad range of pests. We previously demonstrated under laboratory conditions that foliar applications of the defense elicitors Actigard (acibenzolar-S-methyl)...

  8. Impacts of the Psyllid Arytinnis hakani (Homoptera: Psyllidae) on Invasive French Broom in Relation to Plant Size and Psyllid Density.

    Science.gov (United States)

    Hogg, Brian N; Moran, Patrick J; Smith, Lincoln

    2017-06-01

    The impacts of weed biological control agents may vary with plant ontogeny. As plants grow, structural and chemical changes can alter plant resistance, which may reduce herbivory via chemical or structural defenses, and plant tolerance, which may enable plants to maintain fitness despite attack. Resistance and tolerance generally increase as plants grow. Nonetheless, prerelease tests of agent efficacy often overlook plant ontogeny. Here, we assess the performance and impacts of a candidate biocontrol agent, the psyllid Arytinnis hakani (Loginova), in relation to the age of its host plant, the invasive shrub French broom, Genista monspessulana. We also examined whether the psyllid can consistently kill plants when its densities are sufficiently high. Survival of psyllids to adulthood and the timing of adult emergence did not differ between plant sizes, indicating that performance of nymphs was not influenced by plant size. However, adult psyllid survival was reduced on small plants, suggesting that nymphs and adults responded differently to ontogenetic changes in plant quality. Psyllids affected the growth of small and large plants similarly; all measured plant growth parameters were lower in the presence of psyllids regardless of plant size. In a separate experiment, effects on plant survival depended on psyllid density, as higher realized densities of ∼9 psyllids per cm stem length were necessary to consistently kill plants. Thus, results suggest that the psyllid would be equally effective on a range of plant sizes, particularly at high densities, and show the potential of the psyllid to help control French broom in California. Published by Oxford University Press on behalf of Entomological Society of America 2017. This work is written by US Government employees and is in the public domain in the US.

  9. Repellency of a kaolin particle film to potato psyllid, Bactericera cockerelli (Hemiptera: Psyllidae), on tomato under laboratory and field conditions.

    Science.gov (United States)

    Peng, Linian; Trumble, John T; Munyaneza, Joseph E; Liu, Tong-Xian

    2011-07-01

    The potato psyllid, Bactericera cockerelli, is a vector of Candidatus Liberibacter solanacearum, causing several diseases in solanaceous crops. Laboratory and field no-choice and choice experiments were conducted to evaluate the repellency of kaolin particle film on adults of B. cockerelli on tomato plants that had been sprayed with kaolin particle film on the upper surface only, on the lower surface only and on both leaf surfaces. In no-choice tests in the laboratory, the numbers of adults on leaves were not different between the kaolin particle film and the water control, regardless of which leaf surface(s) were treated, but numbers of eggs were lower on the leaves treated with kaolin particle film than on those treated with water. In choice tests on plants treated with water/plants treated with kaolin particle film at ratios of 1:1, 6:3 or 8:1, fewer adults and eggs were found on the leaves treated with kaolin particle film than on leaves treated with water. Under field conditions, in caged no-choice or choice tests, fewer adults, eggs and nymphs were found on plants treated with kaolin particle film than on plants treated with water. In an uncaged test under field conditions, plants sprayed with kaolin particle film had fewer psyllids than those sprayed with water. Even though potato psyllid adults could land on plants treated with kaolin particle film when no choice was given, fewer eggs were laid. When given a choice, the psyllids avoided plants treated with kaolin particle film under laboratory and field conditions. Kaolin particle film treatment may be a useful alternative for management of potato psyllids under field conditions. Copyright © 2011 Society of Chemical Industry.

  10. Repellency of a kaolin particle film to potato payllid, Bactericera cockerelli (Hemiptera: Psyllidae) on tomato under laboratory and field conditions

    Science.gov (United States)

    The potato psyllid, Bactericera cockerelli, is a vector of “Candidatus Liberibacter solanacearum,” the bacterium causing several diseases in solanaceous crops. Laboratory and field no-choice and choice experiments were conducted to evaluate repellency of kaolin particle film on adults of B. cockerel...

  11. Volatile profiles of young leaves of Rutaceae spp. varying in susceptibility to the Asian citrus psyllid,(Hemiptera: Psyllidae)

    Science.gov (United States)

    Plant volatiles were identified from six species in the family Rutaceae. These species had varying degrees of susceptibility to the Asian citrus psyllid as determined by direct counts of life stages. Using a push system involving charcoal-filtered humidified air, volatiles were adsorbed on SuperQ pa...

  12. Synthetic blends of volatile, phytopathogen-induced odorants can be used to manipulate vector behavior.

    Directory of Open Access Journals (Sweden)

    Alexander eAksenov

    2014-12-01

    Full Text Available Volatile organic compounds (VOCs are emitted from all plants and these VOCs are important means of communication between plants and insects. It has been documented that pathogen infections alter VOC profiles rendering infected plants more attractive to specific vectors transmitting these pathogens than uninfected plants, thus potentially aiding in pathogen propagation. Mimicking these chemical cues might enable insect attraction away from the plant or disruption of host finding behavior of the vector. However, the practical implications have not been fully explored. We used citrus, Diaphorina citri and huanglongbing (HLB as a model host-vector-disease system because HLB threatens citrus production worldwide and is similar to other critical diseases of food crops, such as Zebra Chip affecting potato. We formulated a synthetic chemical blend using selected HLB-specific biomarker compounds, and tested the blend with the Attenu assay system for chemosensory proteins. The Attenu assay system is a procedure that identifies interactions between insect chemosensory proteins and their ligands. We found that an equimolar mixture of compounds mimicking the volatile profile of HLB-infected citrus bound chemosensory proteins. Further investigation of this blend in laboratory behavioral assays resulted in development of a synthetic lure that was more attractive to D. citri than natural citrus tree volatiles. This strategy could provide a new route to produce chemical lures for vector population control for a variety of plant and/or animal systems and it may result in the development of a practical lure for monitoring vectors of disease, such as D. citri.

  13. The community composition and temporal dynamic of canopy arthropods on Citrus in Nanchang, Jiangxi Province%江西南昌柑橘冠层节肢动物群落组成及时序动态研究

    Institute of Scientific and Technical Information of China (English)

    李小珍; 王建国; 肖海军; 刘伟; 鲍涵涵; 廖琪

    2013-01-01

    采用叶面观察和诱捕相结合的方法调查了江西南昌柑橘冠层节肢动物群落.共调查到节肢动物数量144037头,分属25种.其中,高密度种类有2种即柑橘全爪螨和矢尖蚧;常见种类有13种,包括桔小实蝇、柑橘花蕾蛆、柑橘木虱和中华草蛉等;其它为低密度种类.在群落结构指数的变化过程中,分别于5月下旬和9月下旬出现Shannon-Wiener多样性指数极大值和Simpson优势度指数极小值.对高密度种类和部分常见种类的种群动态进行了分析,表明柑橘全爪螨在5月下旬至6月上旬发生严重,矢尖蚧则于7~8月发生严重;柑橘花蕾蛆幼虫和柑橘木虱发生动态相似,均仅于柑橘春梢阶段出现最高密度;桔小实蝇成虫则在9月下旬出现密度高峰;中华草蛉的发生动态与其主要捕食对象一柑橘全爪螨的相似.研究结果初步明确南昌柑橘冠层节肢动物的重要成分及重要类群的时序动态,为该地区柑橘重要害虫的防治与控制,天敌昆虫的保护与利用提供基础信息.%The composition and structure of the arthropod communities in orange orchard were investigated in Nanchang,Jiangxi Province,using the leaf observational method and the trapping technique from April to December 2012.A total of 144037 individuals,belonging to 25 species,were founded in our investigation.Among them,the high density species included with Panonychus citri MeGregor and Unaspis yanonensis (Kuwana),13 kinds of common species including Diaphorina Citri (Kuwayama),Bactrocera dorsalis Hendel,Contarinia citri Barnes,Chrysoperla sinica (Tjeder) and so on,and others were the low density species.During the change of arthropod community indexes,Shannon -Wiener diversity index reached their maximum values,and Simpson dominance index appeared their minimum values in late May and late September,respectively.Subsequently,we described the population dynamics of high density species and some common species

  14. Lethal and Sublethal Effects of Insecticides Used on Citrus, on the Ectoparasitoid Tamarixia radiata.

    Science.gov (United States)

    Beloti, Vitor Hugo; Alves, Gustavo Rodrigues; Araújo, Diogo Feliciano Dias; Picoli, Mateus Manara; Moral, Rafael de Andrade; Demétrio, Clarice Garcia Borges; Yamamoto, Pedro Takao

    2015-01-01

    Huanglongbing (HLB) is a disease associated with the bacteria "Candidatus Liberibacter spp." and has been devastating citrus orchards around the world. Its management involves control of the insect vector, the Asian citrus psyllid Diaphorina citri Kuwayama. However, the indiscriminate use of chemicals has caused pest outbreaks and eliminated the natural enemies of the vector, such as the parasitoid Tamarixia radiata (Waterston), the main agent for biological control of D. citri. This study assessed the lethal and sublethal effects of insecticides recommended for integrated production of citrus on the parasitoid T. radiata. When adult parasitoids were exposed to residues of 25 insecticides, 20% of them, i.e., gamma-cyhalothrin, etofenprox, azadirachtin, tebufenozide and pyriproxyfen, were considered as harmless (Class 1), 12% as slightly harmful (Class 2), 12% as moderately harmful (Class 3) and 56% as harmful (Class 4), according to the classification proposed by the IOBC/WPRS. Afterward, 14 insecticides (5 harmless and 9 harmful) were sprayed on the parasitoid pupae. Of the 14 insecticides tested, only the organophosphates dimethoate and chlorpyrifos affected the parasitoid emergence. The effects of insecticides on the parasitism capacity of adults exposed to residues of azadirachtin, etofenprox, gamma-cyhalothrin, pyriproxyfen and tebufenozide (harmless) were also evaluated. Tebufenozide and gamma-cyhalothrin affected the parasitism of the F0 generation, but did not affect the emergence of the F1 and F2 generations. Therefore, for an effective IPM program, selective insecticides or harmful pesticides to adult parasitoids could be used in the field, provided that the adults do not occur naturally and the chemical applications do not coincide with parasitoid releases.

  15. Differential effects of exposure to ambient vanilla and citris aromas on mood, arousal and food choice

    NARCIS (Netherlands)

    Wijk, de R.A.; Zijlstra, S.

    2012-01-01

    Background Aromas have been associated with physiological, psychological affective and behavioral effects. We tested whether effects of low-level exposure to two ambient food-related aromas (citrus and vanilla) could be measured with small numbers of subjects, low-cost physiological sensors and semi

  16. Temporal variation in sex allocation in the mealybug Planococcus citri : Adaptation, constraint, or both?

    NARCIS (Netherlands)

    Ross, Laura; Langenhof, Minke B. W.; Pen, Ido; Shuker, David M.

    2012-01-01

    Sex ratio theory has been very successful in predicting under which circumstances parents should bias their investment towards a particular offspring sex. However, most examples of adaptive sex ratio bias come from species with well-defined mating systems and sex determining mechanisms, while in man

  17. Parallel evolution of cytochrome b mediated bifenazate resistance in the citrus red mite Panonychus citri

    NARCIS (Netherlands)

    Van Leeuwen, T.; Van Nieuwenhuyse, P.; Vanholme, B.; Dermauw, W.; Nauen, R.; Tirry, L.

    2011-01-01

    Bifenazate is a recently developed acaricide that is mainly used to control spider mites on a variety of crops. Although first thought to be a neurotoxin, genetic evidence obtained from bifenazate resistant Tetranychus urticae strains suggested an alternative mode of action as a Qo pocket inhibitor

  18. Molecular evidence of polyandry in the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae.

    Directory of Open Access Journals (Sweden)

    Sofia G Seabra

    Full Text Available The occurrence of polyandry in Planococcuscitri, presumed by earlier observations of mating behavior, was confirmed using microsatellite genotyping of pools of over 400 eggs resulting from controlled crosses of one female with two males. The genetic contribution of both mated males was confirmed in 13 out of 43 crosses. In three crosses it was possible to determine that only the first male fertilized the eggs, which may be due to sperm competition or unviable sperm supply. The microsatellite analysis also allowed the confirmation of aspects of the chromosomal inheritance detected previously in cytogenetic studies in Planococcuscitri, namely that only one of the alleles is transmitted by the male, indicating that the males are functionally haploid, supporting the observation of Paternal Genome Elimination (PGE in these insects.

  19. Molecular evidence of polyandry in the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae).

    Science.gov (United States)

    Seabra, Sofia G; Brás, Patricia G; Zina, Vera; Borges da Silva, Elsa; Rebelo, Maria Teresa; Figueiredo, Elisabete; Mendel, Zvi; Paulo, Octávio S; Franco, José Carlos

    2013-01-01

    The occurrence of polyandry in Planococcuscitri, presumed by earlier observations of mating behavior, was confirmed using microsatellite genotyping of pools of over 400 eggs resulting from controlled crosses of one female with two males. The genetic contribution of both mated males was confirmed in 13 out of 43 crosses. In three crosses it was possible to determine that only the first male fertilized the eggs, which may be due to sperm competition or unviable sperm supply. The microsatellite analysis also allowed the confirmation of aspects of the chromosomal inheritance detected previously in cytogenetic studies in Planococcuscitri, namely that only one of the alleles is transmitted by the male, indicating that the males are functionally haploid, supporting the observation of Paternal Genome Elimination (PGE) in these insects.

  20. Acción inhibitoria de una cepa de Zymomonas mobilis mobilis aislada de caña de azúcar sobre Xanthomonas citri subsp. citri, agente causal de la cancrosis de los cítricos Inhibition of Xanthomonas citri subsp. citri, causal agent of citrus canker, by a strain of Zymomonas mobilis mobilis isolated from sugarcane

    National Research Council Canada - National Science Library

    María E. Romero; C. Jacqueline Ramallo; L. Daniel Ploper

    2008-01-01

    ... patógenos para el hombre, animales y plantas. Un problema importante a resolver en los tratamientos con antimicrobianos, es el desarrollo de resistencia a compuestos empleados actualmente, no siendo las bacterias fitopatógenas una excepción...

  1. Acción inhibitoria de una cepa de Zymomonas mobilis mobilis aislada de caña de azúcar sobre Xanthomonas citri subsp. citri, agente causal de la cancrosis de los cítricos Inhibition of Xanthomonas citri subsp. citri, causal agent of citrus canker, by a strain of Zymomonas mobilis mobilis isolated from sugarcane

    OpenAIRE

    María E. Romero; C. Jacqueline Ramallo; L. Daniel Ploper

    2008-01-01

    Zymomonas mobilis mobilis (Zm) produce factores antimicrobianos que actúan sobre un amplio espectro de microorganismos patógenos para el hombre, animales y plantas. Un problema importante a resolver en los tratamientos con antimicrobianos, es el desarrollo de resistencia a compuestos empleados actualmente, no siendo las bacterias fitopatógenas una excepción. En el presente trabajo se realizaron ensayos de antagonismo con células (pruebas de estrías cruzadas) y sobrenadantes concentrados (Sc) ...

  2. [First record of Zoophthora radicans (Entomophthorales: Entomophthoraceae) on adults of the Paraguay tea psyllid, Gyropsylla spegazziniana Lizer & Trelles (Hemiptera: Psyllidae), in Brazil].

    Science.gov (United States)

    Alves, Luis F A; Leite, Luis G; Oliveira, Daian G P de

    2009-01-01

    We report the first occurrence of Zoophthora radicans infecting adults of Gyropsylla spegazziniana Lizer & Trelles in a commercial Paraguay tea plantation (Ilex paraguariensis), in Cascavel, PR, Brazil. The fungus prevalence was high (90% of mortality), considered a natural epizooty.

  3. Danos causados por Ctenarytaina spatulata Taylor, 1977 (Hemiptera: Psyllidae em Eucalyptus grandis Hill. ex Maiden. Eucalyptus grandis Hill.ex. Maiden injuries caused by Ctenarytaina spatulata Taylor.

    Directory of Open Access Journals (Sweden)

    Dalva Luiz de Queiroz Santana

    2011-03-01

    Full Text Available Os danos de Ctenarytaina spatulata Taylor, 1977 foram caracterizados em observações realizadas em campo,  laboratório e casa de vegetação. Os primeiros danos de C. spatulata em Eucalyptus grandis são provocados  pela postura, ficando uma pequena mancha preta, onde o ovo é depositado, que evolui para manchas maiores,  podendo levar a seca da gema. Os espécimens eliminam grandes quantidades de líquidos ricos em  açúcares “honeydew”, que se acumulam sobre folhas e  ponteiros, propiciando a proliferação de fungos formadores de fumagina e fungos fitopatogênicos. As  plantas atacadas ficam com as folhas deformadas,  menores que as normais, encarquilhadas, com falhas  entre as nervuras e com fumagina.  Conseqüentemente, a capacidade fotossintética  diminui. Com os danos provocados pelas picadas,  posturas, acúmulo de fumagina, aparecimento de  fungos oportunistas e a morte dos brotos, a planta  perde a dominância apical, ocorrendo uma  superbrotação no local. O ataque de C. spatulata causa uma diminuição do diâmetro das plantas e encurtamento dos internódios, ocasionando uma maior  fragilidade na área onde há esta formação. Cortes  anatômicos do caule, infestado com C. spatulata, mostram que a parte interna do lenho, na região onde  houve um superbrotamento, apresenta grande  concentração de fibras gelatinosas, que são menos  lignificadas. Estas fibras conferem menor resistência ao  caule das plantas, tornando-o frágil e susceptível a  quebra em função de ventos ou mesmo pelo próprio  peso. Além disto, as plantas atacadas apresentaram  modificações anatômicas quanto ao arranjo dos poros e  maior espessamento da casca.
    The injuries of Ctenarytaina spatulata Taylor, 1977, were characterized through observations in field,  laboratory and greenhouse. The first injuries of C. spatulata in Eucalyptus grandis are caused during  oviposition. At the point where the egg is inserted, a   small black spot appears, which may grow larger, causing the bud death. The insect eliminates large  amount of honeydew, accumulating on leaves and  apical parts, which starts the proliferation of sooty mold,  and fitopathogenic fungi. Leaves of the attacked  lants become deformed, smaller than the  normal ones, distorted, or with flaws among the ribs and sooty mold reducing the photosynthetic capacity.  Due to damages provoked by the bites, oviposition,  sooty mold accumulation and other fungi, the sprout  die and the plant loses the apical dominance, which  induces an over sprouting. In addition, the attack of C. spatulata causes a decrease of diameter growth and the formation of shorter internodes, resulting in a  higher fragile area. Anatomical cuts of the seedlings,  infested with C. spatulata where there was an over  sprouting, presented areas with larger number of  gelatinous fibers, less  lignified. These fibers are less  resistant, turning the stem more fragile and susceptible  o breaking due to wind or even its own weight. Besides, the attacked  plants presented anatomical modifications in pores arrangement and thicker bark.

  4. [Effect of the length of the stay of the male on the reproduction of the olive psylla Euphyllura olivina Costa (Homoptera, Psyllidae)].

    Science.gov (United States)

    Taktak, A

    1984-01-01

    We have studied, in Euphyllura olivina Costa, 1839, the incidence that the absence or presence of males can have on the fecundity of females their longevity, the duration of spawning and other biotic parameters... So we have pointed out that: In all the cases, from beginning of experimental infestations of olive--tree sprouts to the beginning of eggs releasing, there is a preovipositional time of very variable duration. Virgins, isolated from imago emergence, have a trifling spawning and the eggs cannot have parthenogenetic development. Fecundity of a female by one male induces important spawnings. These increase with more and more prolonged cohabitation (2, 4, 8 and 16 days stay). When one female is in the presence of two males we have noted a very hight variability of fecundity of the female. However it appears that the most important spawnings occur when the two males and the female are left together during 8 days.

  5. Protective effects of egg stalk of Paratrioza sinica (Hemiptera: Psyllidae) at various angles and spacing against three predaceous coccinellids, Harmonia axyridis, Coccinella septempunctata and Hippodamia variegata (Coleoptera: Coccinellidae).

    Science.gov (United States)

    Wu, Pengxiang; Ma, Baoxu; Yan, Shuo; Xu, Jing; He, Jia; Zhang, Rong; Zhang, Runzhi

    2017-08-26

    Paratrioza sinica, is a major pest of wolfberries. Coccinellids could effectively control various developmental stages of P. sinica damage except the stalked egg. To analyze the protective role of egg stalks against predaceous coccinellids, Harmonia axyridis, Coccinella septempunctata and Hippodamia variegata, we studied the functional responses and effects of two potential factors: the angle between egg stalk and leaf plane, and the spacing between egg stalks. The searching rate, handling time and theoretical maximum egg consumption of H. variegata was optimal among 3 ladybug species. The egg consumption by coccinellids were maximum and minimum at 0° and 90°, respectively. Average reduction rates from 0° to 90° of egg consumed by larvae of coccinellids and H. variegata were significantly lower compared to adults and other 2 species, respectively. Optimal spacing of egg consumption varied with different predator species and their developmental stages, which were nearly close to the body lengths of predators. Egg stalk was served as a physical protection against the predators. Selective advantage of egg stalk is a facilitator in protection against predators during evolution, which needs more attention. Reasonable selection of predator and irrigation strategy may exhibit positive control performance. This article is protected by copyright. All rights reserved.

  6. Influence of Temperature, Humidity, and Plant Terpenoid Profiles on Life History Characteristics of Boreioglycaspis melaleucae (Hemiptera: Psyllidae), a Biological Control Agent of the Invasive Tree Melaleuca quinquenervia

    Science.gov (United States)

    Although the introduced weed biological control agent Boreioglycaspis melaleucae Moore is widely established among stands of its host Melaleuca quinquenervia (Cav.) Blake (Myrtaceae) in south Florida, it’s population densities decline markedly during summer months. We investigated the hypothesis tha...

  7. Mosquitocidal carbamates with low toxicity to agricultural pests: an advantageous property for insecticide resistance management.

    Science.gov (United States)

    Swale, Daniel R; Carlier, Paul R; Hartsel, Joshua A; Ma, Ming; Bloomquist, Jeffrey R

    2015-08-01

    Insecticide resistance in the malaria mosquito Anopheles gambiae is well documented, and widespread agricultural use of pyrethroids may exacerbate development of resistance when pyrethroids are used in vector control. We have developed carbamate anticholinesterases that possess a high degree of An. gambiae:human selectivity for enzyme inhibition. The purpose of this study was to assess the spectrum of activity of these carbamates against other mosquitoes and agricultural pests. Experimental carbamates were potent inhibitors of mosquito acetylcholinesterases, with IC50 values in the nanomolar range. Similar potencies were observed for Musca domestica and Drosophila melanogaster enzymes. Although meta-substituted carbamates were potent inhibitors, two ortho-substituted carbamates displayed poor enzyme inhibition (IC50 ≥ 10(-6)  M) in honey bee (Apis mellifera), Asian citrus psyllid (Diaphorina citri) and lepidopteran agricultural pests (Plutella xylostella and Ostrinia nubilalis). Enzyme inhibition results were confirmed by toxicity studies in caterpillars, where the new carbamates were 2-3-fold less toxic than propoxur and up to tenfold less active than bendiocarb, indicating little utility of these compounds for crop protection. The experimental carbamates were broadly active against mosquito species but not agricultural pests, which should mitigate selection for mosquito insecticide resistance by reducing agricultural uses of these compounds. © 2014 Society of Chemical Industry. © 2014 Society of Chemical Industry.

  8. Asian Citrus Psyllid RNAi Pathway – RNAi evidence

    Science.gov (United States)

    Taning, Clauvis N. T.; Andrade, Eduardo C.; Hunter, Wayne B.; Christiaens, Olivier; Smagghe, Guy

    2016-01-01

    Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if the RNAi machinery could be exploited to develop a management strategy for this pest. Analyses of RNAi-related genes in the Asian citrus psyllid genome showed an absence of sequences encoding R2D2, a dsRNA-binding protein that functions as a cofactor of Dicer-2 in Drosophila. Nevertheless, bioassays using an in Planta System showed that the Asian citrus psyllid was very sensitive to ingested dsRNA, demonstrating a strong RNAi response. A small dose of dsRNA administered through a citrus flush was enough to trigger the RNAi mechanism, causing significant suppression of the targeted transcript, and increased psyllid mortality. This study provides evidence of a functional RNAi machinery, which could be further exploited to develop RNAi based management strategies for the control of the Asian citrus psyllid. PMID:27901078

  9. Phenology of Asian citrus psyllid (Hemiptera: Liviidae) and associated parasitoids on two species of Citrus, kinnow mandarin and sweet orange, in Punjab Pakistan.

    Science.gov (United States)

    Khan, Shouket Zaman; Arif, Muhammad Jalal; Hoddle, Christina D; Hoddle, Mark S

    2014-10-01

    The population phenology of Asian citrus psyllid, Diaphorina citri Kuwayama, was monitored weekly for 110 wk on two species of Citrus, kinnow mandarin and sweet orange, at two different research sites in Faisalabad, Punjab Pakistan. Citrus flush growth patterns were monitored and natural enemy surveys were conducted weekly. Flush patterns were similar for kinnow and sweet orange. However, flush on sweet orange was consistently more heavily infested with Asian citrus psyllid than kinnow flush; densities of Asian citrus psyllid eggs, nymphs, and adults were higher on sweet orange when compared with kinnow. When measured in terms of mean cumulative insect or Asian citrus psyllid days, eggs, nymphs, and adults were significantly higher on sweet orange than kinnow. Two parasitoids were recorded attacking Asian citrus psyllid nymphs, Tamarixia radiata (Waterston) and Diaphorencyrtus aligarhensis (Shafee, Alam and Agarwal). The dominant parasitoid species attacking Asian citrus psyllid nymphs on kinnow and sweet orange was T. radiata, with parasitism averaging 26%. D. aligarhensis parasitism averaged 17%. Generalist predators such as coccinellids and chrysopids were collected infrequently and were likely not important natural enemies at these study sites. Immature spiders, in particular, salticids and yellow sac spiders, were common and may be important predators of all Asian citrus psyllid life stages. Low year round Asian citrus psyllid densities on kinnow and possibly high summer temperatures, may, in part, contribute to the success of this cultivar in Punjab where Candidatus Liberibacter asiaticus, the putative causative agent of huanglongbing, a debilitating citrus disease, is widespread and vectored by Asian citrus psyllid.

  10. Transmission and Propagation of 'Candidatus Liberibacter asiaticus' by Grafting with Individual Citrus Leaves.

    Science.gov (United States)

    Hilf, Mark E; Lewis, Reid S

    2016-05-01

    Huanglongbing (HLB) is a chronic, progressive decline disease in citrus associated with a systemic infection by the bacterium 'Candidatus Liberibacter asiaticus'. Transmission of the bacterium in the field is by the Asian citrus psyllid, Diaphorina citri Kuwayama. Experimental propagation of 'Ca. L. asiaticus' is done primarily by grafting pieces of bud wood from an infected plant. To produce a small-scale model system for investigation of pathogen biology, we investigated grafting single leaves from infected citrus plants as sources of inoculum for propagation of the bacterium. In total, 162 plants ranging in age from 3 to 18 months were grafted. Grafting with intact asymptomatic and HLB-symptomatic leaves resulted in 61 of 78 (78%) and 35 of 41 (85%) of the plants infected with 'Ca. L. asiaticus', respectively. Inoculum consisting of the leaf petiole only or only an inoculum tissue remnant under the bark of the receptor tree resulted in 6 of 12 (50%) and 7 of 31 (23%) infected trees, respectively. Real-time polymerase chain reaction (qPCR) assays verified the infection in plants, a majority of which developed the foliar blotchy mottle symptom considered diagnostic for HLB, while some plants also displayed the stunted, chlorotic shoots for which the disease is named. The qPCR data together with the symptoms displayed demonstrated that individual leaves from infected trees can serve as effective inoculum sources for transmission and propagation of 'Ca. L. asiaticus' via grafting.

  11. Development of rapid, sensitive and non-radioactive tissue-blot diagnostic method for the detection of citrus greening.

    Science.gov (United States)

    Nageswara-Rao, Madhugiri; Miyata, Shin-Ichi; Ghosh, Dilip; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame

    2013-01-01

    Citrus huanglongbing (HLB or citrus greening) is one of the most devastating diseases of citrus worldwide. The disease is caused by Gram-negative, phloem-limited α-proteobacterium, 'Candidatus Liberibacter asiaticus', vectored by the psyllid, Diaphorina citri Kuwayama. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection and non-uniform distribution within the tree makes the detection of the pathogen very difficult. Efficient management of HLB disease requires rapid and sensitive detection early in the infection followed by eradication of the source of pathogen and the vector. The polymerase chain reaction (PCR) based method is most commonly employed for screening the infected/suspected HLB plants and psyllids. This is time consuming, cumbersome and not practical for screening large number of samples in the field. To overcome this, we developed a simple, sensitive, non-radioactive, tissue-blot diagnostic method for early detection and screening of HLB disease. Digoxigenin labeled molecular probes specific to 'Ca. L. asiaticus' nucleotide sequences have been developed and used for the detection of the pathogen of the HLB disease. The copy number of the target genes was also assessed using real-time PCR experiments and the optimized real-time PCR protocol allowed positive 'Ca. L. asiaticus' detection in citrus samples infected with 'Ca. L. asiaticus' bacterium.

  12. Comparison of Potato and Asian Citrus Psyllid Adult and Nymph Transcriptomes Identified Vector Transcripts with Potential Involvement in Circulative, Propagative Liberibacter Transmission

    Directory of Open Access Journals (Sweden)

    Tonja W. Fisher

    2014-11-01

    Full Text Available The potato psyllid (PoP Bactericera cockerelli (Sulc and Asian citrus psyllid (ACP Diaphorina citri Kuwayama are the insect vectors of the fastidious plant pathogen, Candidatus Liberibacter solanacearum (CLso and Ca. L. asiaticus (CLas, respectively. CLso causes Zebra chip disease of potato and vein-greening in solanaceous species, whereas, CLas causes citrus greening disease. The reliance on insecticides for vector management to reduce pathogen transmission has increased interest in alternative approaches, including RNA interference to abate expression of genes essential for psyllid-mediated Ca. Liberibacter transmission. To identify genes with significantly altered expression at different life stages and conditions of CLso/CLas infection, cDNA libraries were constructed for CLso-infected and -uninfected PoP adults and nymphal instars. Illumina sequencing produced 199,081,451 reads that were assembled into 82,224 unique transcripts. PoP and the analogous transcripts from ACP adult and nymphs reported elsewhere were annotated, organized into functional gene groups using the Gene Ontology classification system, and analyzed for differential in silico expression. Expression profiles revealed vector life stage differences and differential gene expression associated with Liberibacter infection of the psyllid host, including invasion, immune system modulation, nutrition, and development.

  13. A review of techniques for detecting Huanglongbing (greening) in citrus.

    Science.gov (United States)

    Arredondo Valdés, Roberto; Delgado Ortiz, Juan C; Beltrán Beache, Mariana; Anguiano Cabello, Julia; Cerna Chávez, Ernesto; Rodríguez Pagaza, Yolanda; Ochoa Fuentes, Yisa M

    2016-10-01

    Huanglongbing (HLB) is the most destructive disease of citrus worldwide. Monitoring of health and detection of diseases in trees is critical for sustainable agriculture. HLB symptoms are virtually the same wherever the disease occurs. The disease is caused by Candidatus Liberibacter spp., vectored by the psyllids Diaphorina citri Kuwayama and Trioza erytreae. Electron microscopy was the first technique used for HLB detection. Nowadays, scientists are working on the development of new techniques for a rapid HLB detection, as there is no sensor commercially accessible for real-time assessment of health conditions in trees. Currently, the most widely used mechanism for monitoring HLB is exploration, which is an expensive, labor-intensive, and time-consuming process. Molecular techniques such as polymerase chain reaction are used for the identification of HLB disease, which requires detailed sampling and processing procedures. Furthermore, investigations are ongoing in spectroscopic and imaging techniques, profiling of plant volatile organic compounds, and isothermal amplification. This study recognizes the need for developing a rapid, cost-effective, and reliable health-monitoring sensor that would facilitate advancements in HLB disease detection. This paper compares the benefits and limitations of these potential methods for HLB detection.

  14. PHLOEM PROMOTERS IN TRANSGENIC SWEET ORANGE ARE DIFFERENTIALLY TRIGGERED BY Candidatus Liberibacter asiaticus

    Directory of Open Access Journals (Sweden)

    LUZIA YURIKO MIYATA

    Full Text Available ABSTRACT The use of promoters preferentially expressed in specific plant tissues is a desirable strategy to search for resistance for pathogens that colonize these tissues. The bacterium Candidatus Liberibacter asiaticus (Las, associated with huanglongbing disease (HLB of citrus, colonizes phloem vessels. Some promoters, besides conferring tissue-specific expression, can also respond to the presence of the pathogen. The objective of the present study was to verify if the presence of Las could modulate the activation of the phloem-specific promoters AtPP2 (Arabidopsis thaliana phloem protein 2, AtSUC2 (A. thaliana sucrose transporter 2 and CsPP2 ( pCitrus phloemrotein 2, known to be expressed in Citrus sinensis phloem. ‘Hamlin’ sweet orange plants (Citrus sinensis L. Osbeck transformed with the uidA (GUS reporter gene under the control of AtPP2, AtSUC2 and CsPP2 promoters were infected to evaluate the interdependence between transgene expression and the concentration of Las. Plants were inoculated with Las by Diaphorina citri and eighteen months later, bacterial concentration and uidA expression were determined by qPCR and RT-qPCR, respectively. Reporter gene expression driven by AtSUC2 promoter was strongly and positively correlated with Las concentration. Therefore, this promoter combines desirable features of both tissue-specificity and pathogen-inducibility for the production of transgenic plants tolerant to Las.

  15. Metalized polyethylene mulch to repel Asian citrus psyllid, slow spread of huanglongbing and improve growth of new citrus plantings.

    Science.gov (United States)

    Croxton, Scott D; Stansly, Philip A

    2014-02-01

    Greening or huanglongbing (HLB) is a debilitating disease of citrus caused by Candidatus Liberibactor asiaticus and transmitted by the Asian citrus psyllid (ACP), Diaphorina citri. HLB now occurs worldwide in all major citrus growing regions except the Mediterranean and Australia. Management relies principally on insecticidal control of the ACP vector, but is insufficient, even for young trees which are most susceptible to the disease. We tested the ability of metalized polyethylene mulch to repel adult ACP as well as effects on incidence of HLB and early tree growth. Metalized mulch significantly reduced ACP populations and HLB incidence compared to whiteface mulch or bare ground. In addition, metalized mulch, together with the associated drip irrigation and fertigation system, increased soil moisture, reduced weed pressure, and increased tree growth rate. Metalized mulch slows spread of ACP and therefore HLB pressure on young citrus trees. Metalized mulch can thereby augment current control measures for young trees based primarily on systemic insecticides. Additional costs could be compensated for by increased tree growth rate which would shorten time to crop profitability. These advantages make a compelling case for large-scale trials using metalized mulch in young citrus plantings threatened by HLB. © 2013 Society of Chemical Industry.

  16. Asymptomatic spread of huanglongbing and implications for disease control.

    Science.gov (United States)

    Lee, Jo Ann; Halbert, Susan E; Dawson, William O; Robertson, Cecile J; Keesling, James E; Singer, Burton H

    2015-06-16

    Huanglongbing (HLB) is a bacterial infection of citrus trees transmitted by the Asian citrus psyllid Diaphorina citri. Mitigation of HLB has focused on spraying of insecticides to reduce the psyllid population and removal of trees when they first show symptoms of the disease. These interventions have been only marginally effective, because symptoms of HLB do not appear on leaves for months to years after initial infection. Limited knowledge about disease spread during the asymptomatic phase is exemplified by the heretofore unknown length of time from initial infection of newly developing cluster of young leaves, called flush, by adult psyllids until the flush become infectious. We present experimental evidence showing that young flush become infectious within 15 d after receiving an inoculum of Candidatus Liberibacter asiaticus (bacteria). Using this critical fact, we specify a microsimulation model of asymptomatic disease spread and intensity in a grove of citrus trees. We apply a range of psyllid introduction scenarios to show that entire groves can become infected with up to 12,000 psyllids per tree in less than 1 y, before most of the trees show any symptoms. We also show that intervention strategies that reduce the psyllid population by 75% during the flushing periods can delay infection of a full grove, and thereby reduce the amount of insecticide used throughout a year. This result implies that psyllid surveillance and control, using a variety of recently available technologies, should be used from the initial detection of invasion and throughout the asymptomatic period.

  17. Immunogenesity of spesific protein molecular weight 16 KDa (PS16 leaf of siam citrus infected by citrus vein phloem degeneration (CVPD disease

    Directory of Open Access Journals (Sweden)

    Made Sritamin

    2012-02-01

    Full Text Available Citrus Vein Phloem degeneration (CVPD is an important citrus disese, which damaged citrus plantation and causing decrease of citrus production. In Indonesia, the CVPD disease caused by Liberobacter asiaticum bactery and the disease spread out by vectir insect Diaphorina citri and using infected bud in wood grafting. In infected citrus plant, two specific protein molecules with molecular weigt 16 kDa and 66 kDa are found. These protein molecules are not found in healthy citrus plant. The immunogenicity of PS16 accumulated on leaf of citrus plant infected by CVPD is known yet. The research material were leaves of citrus plant infected CVPD, leaves of healthy citrus plant and reagent used these research are for isolation of the total protein leaf of citrus plant, SDS-PAGE electroforesis, electroelution of PS16, ELISA Methods, Dot-Blot Method, anti-PS16 as aprimery antibody and secondary antibody is anti-Rabbit IgG Conjugated AP. The result of the research showed that of PS16 accumulated on leaf of citrus plant infected CVPD has immunogenic character. It is indicated by increase of the titer anti-PS16 after first immunization ang 2nd booster by indirect ELISA method and can be used to induce antibody (anti-PS16 and so showed that positive reaction between PS16 with anti-PS16. It is indicated by purples dark blue on cellulose membrane by Dot Blot method.

  18. Nucleotides, micro- and macro-nutrients, limonoids, flavonoids, and hydroxycinnamates composition in the phloem sap of sweet orange.

    Science.gov (United States)

    Hijaz, Faraj; Manthey, John A; Van der Merwe, Deon; Killiny, Nabil

    2016-06-02

    Currently, the global citrus production is declining due to the spread of Huanglongbing (HLB). HLB, otherwise known as citrus greening, is caused by Candidatus Liberibacter asiaticus (CLas) and is transmitted by the Asian citrus psyllids (ACP), Diaphorina citri Kuwayama. ACP transmits CLas bacterium while feeding on the citrus phloem sap. Multiplication of CLas in the phloem of citrus indicates that the sap contains all the essential nutrients needed for CLas. In this study, we investigated the micro- and macro-nutrients, nucleotides, and others secondary metabolites of phloem sap from pineapple sweet orange. The micro- and macro-nutrients were analyzed using inductively coupled plasma-mass spectroscopy (ICP-MS) and inductively coupled plasma-optical emission spectroscopy (ICP-OES). Nucleotides and other secondary metabolites analysis was accomplished by reversed phase HPLC coupled with UV, fluorescence detection, or negative mode electrospray ionization mass spectrometry (ESI-MS). Calcium (89 mM) was the highest element followed by potassium (38.8 mM) and phosphorous (24 mM). Magnesium and sulfur were also abundant and their concentrations were 15 and 9 mM, respectively. The rest of the elements were found in low amounts (sap.

  19. Changes in Variable Number of Tandem Repeats in 'Candidatus Liberibacter asiaticus' through Insect Transmission.

    Directory of Open Access Journals (Sweden)

    Hiroshi Katoh

    Full Text Available Citrus greening (huanglongbing is the most destructive citrus disease worldwide. The disease is associated with three species of 'Candidatus Liberibacter' among which 'Ca. Liberibacter asiaticus' has the widest distribution. 'Ca. L. asiaticus' is commonly transmitted by a phloem-feeding insect vector, the Asian citrus psyllid Diaphorina citri. A previous study showed that isolates of 'Ca. L. asiaticus' were clearly differentiated by variable number of tandem repeat (VNTR profiles at four loci in the genome. In this study, the VNTR analysis was further validated by assessing the stability of these repeats after multiplication of the pathogen upon host-to-host transmission using a 'Ca. L. asiaticus' strain from Japan. The results showed that some tandem repeats showed detectable changes after insect transmission. To our knowledge, this is the first report to demonstrate that the repeat numbers VNTR 002 and 077 of 'Ca. L. asiaticus' change through psyllid transmission. VNTRs in the recipient plant were apparently unrelated to the growing phase of the vector. In contrast, changes in the number of tandem repeats increased with longer acquisition and inoculation access periods, whereas changes were not observed through psyllid transmission after relatively short acquisition and inoculation access periods, up to 20 and 19 days, respectively.

  20. Primer registro para la Argentina de una especie de chicharrita, Ctenarytaina spatulata (Hemiptera: Psyllidae, en plantaciones de eucalipto en Entre Ríos New record in Argentina of a species of jumping plant-lice, Ctenarytaina spatulata (Hemiptera: Psyllidae, in eucalypt plantations from Entre Ríos

    Directory of Open Access Journals (Sweden)

    Juan P. R. Bouvet

    2008-06-01

    Full Text Available Se menciona por primera vez para Argentina la especie Ctenarytaina spatulata Taylor, asociada a plantaciones de eucalipto. Este nuevo registro se obtuvo en la Provincia de Entre Ríos, Argentina.Ctenarytaina spatulata Taylor, a species associated with eucalypt plantations, is recorded for the first time from Entre Ríos, Argentina.

  1. Unusual Behaviour — Unusual Morphology: Mutualistic Relationships between Ants (Hymenoptera: Formicidae) and Diaphorina enderleini (Hemiptera: Psylloidea), Associated with Vernonia amygdalina (Asteraceae)

    National Research Council Canada - National Science Library

    Désirée Chantal Aléné; Champlain Djiéto-Lordon; Daniel Burckhardt

    2011-01-01

    .... The lack of wax deprives them of the protection common in various free-living psyllids. Consequently the psyllid is involved in an unusual mutualistic interaction with the ants Pheidole megacephala and Crematogaster striatula...

  2. Application of droplet digital PCR for quantitative detection of Spiroplasma citri in comparison with real time PCR

    Science.gov (United States)

    Droplet digital Polymerase chain reaction (ddPCR) is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. It is a promising DNA quantification technology for medical diagnostics but there are only a few reports of its use for plant pat...

  3. Analysis of Transcriptome Differences between Resistant and Susceptible Strains of the Citrus Red Mite Panonychus citri (Acari: Tetranychidae)

    OpenAIRE

    Bin Liu; Gaofei Jiang; Yunfei Zhang; Junli Li; Xiaojiao Li; Jiansu Yue; Fei Chen; Haoqiang Liu; Hongjun Li; Shiping Zhu; Jinjun Wang; Chun Ran

    2011-01-01

    BACKGROUND: The citrus red mite is a worldwide citrus pest and a common sensitizing allergen of asthma and rhinitis. It has developed strong resistance to many registered acaricides, However, the molecular mechanisms of resistance remain unknown. we therefore used next generation sequencing technology to investigate the global transcriptomes between resistant strains and susceptible strains. RESULTS: We obtained 34,159, 30,466 and 32,217 unigenes by assembling the SS reads, RS reads and SS&RS...

  4. Transmission Rates of ‘Ca. Liberibacter asiaticus’ by Asian Citrus Psyllid Are Enhanced by the Presence and Developmental Stage of Citrus Flush.

    Science.gov (United States)

    Hall, David G; Albrecht, Ute; Bowman, Kim D

    2016-04-01

    Asian citrus psyllid (Diaphorina citri Kuwayama) transmits a bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) putatively responsible for a devastating citrus disease known as Asiatic huanglongbing (HLB) (citrus greening disease). The psyllid and disease have invaded many citrus-growing regions including the United States, where the disease is seriously jeopardizing the Florida citrus industry. We recently concluded research that showed CLas transmission rates are increased when citrus flush is present. Flush is any new leaf growth ranging in development from first emergence up until the leaves are fully expanded yet still tender. In an experiment with seedlings of a rootstock cultivar ‘US-942’, a 1-wk infestation of 20 Asian citrus psyllids from an infected colony resulted in 53–60% of seedlings becoming infected when flush was present compared with only 7% when no flush was present. In a second experiment with ‘US-942’, 77–97% of seedlings became infected when flush was present compared with 40% when no flush was present. A similar experiment with ‘Valencia’ sweet orange resulted in 23, 80, and 3% seedlings becoming infected when young, older, or no flush was present, respectively. Young plants are therefore more likely to contract HLB if flush is present, with older flush promoting higher infection rates under the conditions of this study. Based on this finding, healthy citrus should be protected from Asian citrus psyllid infestations throughout a flush. To evaluate germplasm for CLas resistance, inoculations using infected Asian citrus psyllid would best be achieved if flush is present.

  5. 台州柑橘黄龙病防控研究新进展%The Control Research Progress of Liberobacter asiaticum in Taizhou City

    Institute of Scientific and Technical Information of China (English)

    余继华; 张敏荣; 陶健; 卢璐; 杨晓; 汪恩国; 叶志勇

    2015-01-01

    根据十几年对柑橘木虱和柑橘黄龙病发生规律与防控技术研究探讨,明确了柑橘木虱在台州橘林的发生分布规律和抽样技术,分析了柑橘木虱虫株率和带毒率与黄龙病、黄龙病与柑橘产量损失等之间的关系,探讨了柑橘黄龙病预警和柑橘木虱防治指标,建立了不同管理方式下的疫情扩散模型预测橘林经济寿命,提出了柑橘黄龙病综合防控技术和可防可控的理念。%After decades of researches and discussions on occurrence regularity and control technology of citrus psyl id (Diaphorina citri Kuwayana) and Liberobacter asi-aticum, the occurrence regularity and sampling technique of citrus psyl id in orange forests in Taizhou was defined and the connections between the infection rate and carrying rate of citrus psyl id and Liberobacter asiaticum as wel as the correlations between the Liberobacter asiaticum and citrus yield loss were discussed. This paper discussed the warnings of Liberobacter asiaticum and citrus psylid control index, the economic life span of orange forests was then predicted by building the diseases spreading models with different management styles. At last, the paper put forward the comprehensive prevention and control technology and the concept that Liber-obacter asiaticum was preventable and control able.

  6. Economic injury levels for Asian citrus psyllid control in process oranges from mature trees with high incidence of huanglongbing.

    Science.gov (United States)

    Monzo, Cesar; Stansly, Philip A

    2017-01-01

    The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is the key pest of citrus wherever it occurs due to its role as vector of huanglongbing (HLB) also known as citrus greening disease. Insecticidal vector control is considered to be the primary strategy for HLB management and is typically intense owing to the severity of this disease. While this approach slows spread and also decreases severity of HLB once the disease is established, economic viability of increasingly frequent sprays is uncertain. Lacking until now were studies evaluating the optimum frequency of insecticide applications to mature trees during the growing season under conditions of high HLB incidence. We related different degrees of insecticide control with ACP abundance and ultimately, with HLB-associated yield losses in two four-year replicated experiments conducted in commercial groves of mature orange trees under high HLB incidence. Decisions on insecticide applications directed at ACP were made by project managers and confined to designated plots according to experimental design. All operational costs as well as production benefits were taken into account for economic analysis. The relationship between management costs, ACP abundance and HLB-associated economic losses based on current prices for process oranges was used to determine the optimum frequency and timing for insecticide applications during the growing season. Trees under the most intensive insecticidal control harbored fewest ACP resulting in greatest yields. The relationship between vector densities and yield loss was significant but differed between the two test orchards, possibly due to varying initial HLB infection levels, ACP populations or cultivar response. Based on these relationships, treatment thresholds during the growing season were obtained as a function of application costs, juice market prices and ACP densities. A conservative threshold for mature trees with high incidence of HLB would help maintain economic

  7. Combining 'omics and microscopy to visualize interactions between the Asian citrus psyllid vector and the Huanglongbing pathogen Candidatus Liberibacter asiaticus in the insect gut.

    Science.gov (United States)

    Kruse, Angela; Fattah-Hosseini, Somayeh; Saha, Surya; Johnson, Richard; Warwick, EricaRose; Sturgeon, Kasie; Mueller, Lukas; MacCoss, Michael J; Shatters, Robert G; Cilia Heck, Michelle

    2017-01-01

    Huanglongbing, or citrus greening disease, is an economically devastating bacterial disease of citrus. It is associated with infection by the gram-negative bacterium Candidatus Liberibacter asiaticus (CLas). CLas is transmitted by Diaphorina citri, the Asian citrus psyllid (ACP). For insect transmission to occur, CLas must be ingested during feeding on infected phloem sap and cross the gut barrier to gain entry into the insect vector. To investigate the effects of CLas exposure at the gut-pathogen interface, we performed RNAseq and mass spectrometry-based proteomics to analyze the transcriptome and proteome, respectively, of ACP gut tissue. CLas exposure resulted in changes in pathways involving the TCA cycle, iron metabolism, insecticide resistance and the insect's immune system. We identified 83 long non-coding RNAs that are responsive to CLas, two of which appear to be specific to the ACP. Proteomics analysis also enabled us to determine that Wolbachia, a symbiont of the ACP, undergoes proteome regulation when CLas is present. Fluorescent in situ hybridization (FISH) confirmed that Wolbachia and CLas inhabit the same ACP gut cells, but do not co-localize within those cells. Wolbachia cells are prevalent throughout the gut epithelial cell cytoplasm, and Wolbachia titer is more variable in the guts of CLas exposed insects. CLas is detected on the luminal membrane, in puncta within the gut epithelial cell cytoplasm, along actin filaments in the gut visceral muscles, and rarely, in association with gut cell nuclei. Our study provides a snapshot of how the psyllid gut copes with CLas exposure and provides information on pathways and proteins for targeted disruption of CLas-vector interactions at the gut interface.

  8. Chemical control of the Asian citrus psyllid and of huanglongbing disease in citrus.

    Science.gov (United States)

    Boina, Dhana Raj; Bloomquist, Jeffrey R

    2015-06-01

    By 2014, huanglongbing (HLB), the most destructive disease of citrus, and its insect vector, the Asian citrus psyllid (ACP), Diaphorina citri (Kuwayama), became established in all major citrus-growing regions of the world, including the United States, with the exception of California. At present, application of insecticides is the most widely followed option for reducing ACP populations, while application of antibiotics for suppressing HLB disease/symptoms is being practiced in some citrus-growing regions. Application of insecticides during the dormant winter season, along with cultivation of HLB-free seedlings and early detection and removal of symptomatic and asymptomatic trees, has been very effective in managing ACP. Area-wide management of ACP by application of insecticides at low volume in large areas of citrus cultivation has been shown to be effective in managing HLB and reducing management costs. As insecticide resistance is a major problem in sustainable management of ACP, rotation/alternation of insecticides with different chemistries and modes of action needs to be followed. Besides control of the insect vector, use of antibiotics has temporarily suppressed the symptoms of HLB in diseased trees. Recent efforts to discover and screen existing as well as new compounds for their antibiotic and antimicrobial activities have identified some promising molecules for HLB control. There is an urgent need to find a sustainable solution to the HLB menace through chemical control of ACP populations and within HLB-infected trees through the judicious use of labeled insecticides (existing and novel chemistries) and antibiotics in area-wide management programs with due consideration to the insecticide resistance problem. © 2014 Society of Chemical Industry.

  9. Male Psyllids Differentially Learn in the Context of Copulation

    Directory of Open Access Journals (Sweden)

    Dara G. Stockton

    2017-02-01

    Full Text Available In the Asian citrus psyllid, Diaphorina citri Kuwayama, stimulatory cuticular hydrocarbons act as sex pheromone attractants. Male psyllids locate aggregations of females using those olfactory cues, as well as vibrational communication on the plant surface. Although previous research has indicated that learning plays a role in modulating female reproductive behaviors in psyllids, it is unknown whether males similarly use learning to increase the likelihood of copulatory success. We used an olfactometer-based bio-assay to study the effects of experience on male response to female odor. First, we compared male attraction to female odor in virgin and previously mated males. Second, we tested the effect of several modes of experience with a novel odor, vanillin, to determine whether mating, feeding, or general environmental exposure elicited a learned response. We found that male attraction to female odor significantly increased after mating experience. In addition, we found that males learn about odor specifically in the context of mating, rather than feeding or general exposure. Electrophysiological measurements of antennal response to odorants confirmed that mating status did not affect the sensitivity of the peripheral nervous system to volatile stimuli implicating learning at the level of the central nervous system. These results suggest that male response to female odor is not an entirely innate behavior. Males may require mating experience with female conspecifics to develop attraction to those olfactory cues produced by the female and in association with the female’s habitat. This adaptive plasticity may allow males to detect females in an ever-changing environment and promote diversification and further specialization on different host genotypes.

  10. Male Psyllids Differentially Learn in the Context of Copulation

    Science.gov (United States)

    Stockton, Dara G.; Martini, Xavier; Stelinski, Lukasz L.

    2017-01-01

    In the Asian citrus psyllid, Diaphorina citri Kuwayama, stimulatory cuticular hydrocarbons act as sex pheromone attractants. Male psyllids locate aggregations of females using those olfactory cues, as well as vibrational communication on the plant surface. Although previous research has indicated that learning plays a role in modulating female reproductive behaviors in psyllids, it is unknown whether males similarly use learning to increase the likelihood of copulatory success. We used an olfactometer-based bio-assay to study the effects of experience on male response to female odor. First, we compared male attraction to female odor in virgin and previously mated males. Second, we tested the effect of several modes of experience with a novel odor, vanillin, to determine whether mating, feeding, or general environmental exposure elicited a learned response. We found that male attraction to female odor significantly increased after mating experience. In addition, we found that males learn about odor specifically in the context of mating, rather than feeding or general exposure. Electrophysiological measurements of antennal response to odorants confirmed that mating status did not affect the sensitivity of the peripheral nervous system to volatile stimuli implicating learning at the level of the central nervous system. These results suggest that male response to female odor is not an entirely innate behavior. Males may require mating experience with female conspecifics to develop attraction to those olfactory cues produced by the female and in association with the female’s habitat. This adaptive plasticity may allow males to detect females in an ever-changing environment and promote diversification and further specialization on different host genotypes. PMID:28178203

  11. Initial impacts and field validation of host range for Boreioglycaspis melaleucae Moore (Hemiptera: Psyllidae),a biological control agent of the invasive tree Melaleuca quinquenervia (Cav.) Blake (Myrtales: Myrtaceae: Leptosp

    Science.gov (United States)

    Invasion of south Florida wetlands by the Australian paperbark tree (“melaleuca”), Melaleuca quinquenervia (Cav.) S.T. Blake (melaleuca) has caused adverse economic and environmental impacts. The tree’s biological attributes along with favorable ambient biophysical conditions combine to complicate ...

  12. Flutuação populacional de Gyropsylla spegazziniana (Lizer y Trelles (Hemiptera, Psyllidae e de seus inimigos naturais em erva-mate no município de São Mateus do Sul, PR, Brasil Population dynamics of the Gyropsylla spegazziniana (Lizer y Trelles (Hemiptera, Psyllidae and your natural enemies in planted "ervais", in the municipal district of São Mateus do Sul, PR, Brazil

    Directory of Open Access Journals (Sweden)

    Silvia Pereira Leite

    2007-12-01

    Full Text Available G. spegazziniana é considerada uma das principais pragas da cultura da erva mate, porém pouco se conhece sobre sua densidade populacional e de seus inimigos naturais. O objetivo do trabalho foi determinar a dinâmica populacional da praga e de seus inimigos naturais, visando definir o momento adequado para seu controle. A flutuação populacional foi avaliada mediante a instalação de 10 armadilhas Gyrotrap®, em uma área de 0,5 ha, em um erval situado em São Mateus do Sul, Pr, Brasil. Verificou-se a ocorrência de G. spegazziniana durante o ano todo, com um aumento populacional no início e meados da primavera; os picos foram observados entre outubro e abril. Para o levantamento dos inimigos naturais, foram realizadas coletas quinzenais diretamente nas erveiras, utilizando-se um funil de alumínio. Dentre os inimigos naturais associados à cultura, a maioria foram predadores e uma espécie de parasitóide, os quais apresentaram picos populacionais sincronizados com o da praga.Gyropsylla spegazziniana (Lizer & Trelles, 1919 is considered to be one of the principal pests of "erva-mate" (Ilex paraguariensis plantations, but little is known about the density of its population and their natural enemies along the year. The objective of the present research is to study the population dynamics of the pest as well as its natural enemies, aiming to define the best moment for its control. To measure pest population dynamics, ten "Gyrotrap®" traps were installed an area of 0.5 ha of "erva-mate" plantation in São Mateus do Sul, Pr. Brasil. G. spegazziniana was found to be present throughout the year, with population growth highest in the beginning and middle of spring, and peaks between October and April. To survey natural enemies of the pest species, collections were made every two weeks in the plantations, using an aluminum funnel. Predators and a species of parasitoids were the natural enemies associated to the culture. They also showed synchronism with the peak occurrence of the pest.

  13. Gender- and species-specific characteristics of bacteriomes from three psyllid species (Hemiptera: Psylloidae)

    Science.gov (United States)

    Psyllids (Hemiptera: Pyslloidea) harbor bacterial symbionts in specialized organs called bacteriomes. Bacteriomes may be subject to manipulation to control psyllid pests including Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) and Cacopsylla pyricola (Forster) (Hemiptera: Psyllidae) if the bi...

  14. 福建省柑橘黄龙病危害及其流行规律研究%Damage and Epidemics of Citrus Huanglongbing in Fujian Province

    Institute of Scientific and Technical Information of China (English)

    谢钟琛; 李健; 施清; 谢文龙; 杨建榕; 陈跃飞; 钟连生

    2009-01-01

    [目的]全面了解柑橘黄龙病(Citrus Huanglongbing,简称HLB)在福建省的危害分布及其流行情况,有效指导HLB的防控.[方法]对全省48个县(市、区)及245个柑橘主产乡镇HLB虫媒柑橘木虱(Diaphorina citri)分布及HLB的危害进行调查,以PCR(polymerase chain reaction,PCR)方法对采集的斑驳黄化柑橘叶片、红鼻果进行确证性检测,分析HLB的危害流行规律,并应用滑动相关分析筛选影响HLB流行的气象要素.[结果]全省各县及柑橘主产乡镇均可查到柑橘木虱和HLB病株(最北N28°07'.海拔最高880 m),HLB危害程度为平均发病率16.59%,主栽品种间感病性差异显著,排序为宽皮桔>甜橙类>柚类;HLB发病率与果园纬度、海拔、坡度呈显著负相关;在南亚热带气候区的长泰县(N24°07'),HLB田间发病率仅与临近2年春季气候有关,推测HLB潜伏期为6~18个月,同时春梢期日照、幼果期湿度与HLB发病率呈极显著正、负相关,"春旱"为HLB流行性气候;柑橘红鼻果、斑驳黄化叶片的HLB病原Nested-PCR检出率分别为93.8%、73.1%,前者可靠性优于后者,二类症状均可视为HLB的典型症状.[结论]福建境内的柑橘产区均有HLB病原与柑橘木虱的分布.若春季遇严重干旱HLB就有局部爆发的可能.柑橘红鼻果可作HLB田间诊断依据.%[Objective] To effectively control citrus huanglongbing (HLB, yellow shoot disease), the distribution, damage and epidemics of the disease were investigated in Fujian Province. [Method] The distribution of"Candidatus Liberibacter asiaticus", the putative pathogen, the Asian citrus psyllid (Diaphorina citri), the insect vectors, and the incidence of HLB were investigated in 245 citrus-producing towns in 48 counties in Fujian Province. Polymerase chain reaction (PCR) assay was carried out to verify the existence of HLB in the collected blotchy mottle leaves and color inversion fruits to analyze the epidemics of HLB. Sliding correlation

  15. Study on HPLC Fingerprint of Folium Citri Grandis of Tongxian%通贤柚叶HPLC指纹图谱研究

    Institute of Scientific and Technical Information of China (English)

    游元元; 范媛媛; 陈席; 谢仁宗

    2014-01-01

    目的 建立通贤柚叶的HPLC指纹图谱.方法 色谱柱为Sepax Amethyst C18-H(250 mm×4.6 0mm,5μm),流动相为乙腈-0.1%甲酸,梯度洗脱,流速1.0 mL· min-1,检测波长303 nm,柱温25℃,进样量20μL.分析12批通贤柚叶样品,并进行主成分分析.结果 建立了通贤柚叶的HPLC指纹图谱,确定了16个共有色谱峰,采用对照品指认了其中2个峰.主成分分析结果表明,11个色谱峰对其质量控制较为重要.结论 所建立的指纹图谱分析方法特征性强,为通贤柚叶的质量控制提供了科学依据.

  16. The occurrence, prevention and cure of Exocarpium Citri Grandis plant diseases and insect pests%化橘红病虫害的发生与防治

    Institute of Scientific and Technical Information of China (English)

    蔡岳文; 严振; 丘金裕

    2004-01-01

    化橘红为芸香科植物化州柚(Citrus grandis(L)Osbeck var.tomentosa Hort)以未成熟果实的外果皮入药。近几年来,随着基地化、规模化大面积种植,其病虫害的发生及危害程度亦与日俱增,一些重要害虫已严重影响化橘红的生产和产品质量。

  17. Sex allocation in a species with paternal genome elimination : The roles of crowding and female age in the mealybug Planococcus citri

    NARCIS (Netherlands)

    Ross, Laura; Langenhof, Minke B. W.; Pen, Ido; Beukeboom, Leo W.; West, Stuart A.; Shuker, David M.

    2010-01-01

    Background: In species with paternal genome elimination, both sexes are diploid. However, in males the chromosomes inherited from the father are deactivated during early development and eliminated from the germ line. Sex allocation theory predicts that, all else being equal, females should bias thei

  18. Identification of Bacterial Plant Pathogens Using Multilocus Polymerase Chain Reaction/Electrospray Ionization-Mass Spectrometry

    Science.gov (United States)

    2008-01-01

    seedlings (sweet orange ) were inoculated with X. citri subsp. citri (also known as X. axonopodis pv. citri). Cabbage plants were inoculated with X...caryophylli PC113; ATCC25418 Carnation ; USA 1 Burkholderia gladioli pv. gladioli FC-368; PM107; ATCC10248 Gladiolus; USA 1 Clavibacter michiganensis...alfalfae pv. citrumelosis No call – Citrus inoculated with Xanthomonas alfalfae pv. citrumelosis No call – Healthy citrus (sweet orange ) No call

  19. 宜昌市柑桔病虫发生新动态及防治策略%Dynamics and control strategy of citrus pests in Yichang City

    Institute of Scientific and Technical Information of China (English)

    张国宝

    2000-01-01

    本文着重介绍了近几年来,宜昌市柑桔红蜘蛛(Panxonychus citri McGregor)、矢尖蚧(Unaspis yanonensis Kuwana)、蚜虫(Aphis citricidus Kirkaldy)、大实蝇(Tetradacus citri Chen)、吉丁虫(Agrilus citri Matsumura)、疮痂病(Elsinoe fawcetti But.et Jenk)、煤烟病(Capncdium citri Berk.et Desm)等发生新动态,通过研究分析其新动态,针对性地提出了今后柑桔病虫的综防策略.

  20. Stylet penetration of Cacopsylla pyri; an electrical penetration graph (EPG) study

    NARCIS (Netherlands)

    Civolani, S.; Leis, M.; Grandi, G.; Garzo, E.; Pasqualini, E.; Musacchi, S.; Chicca, M.; Castaldelli, G.; Rossie, M.; Tjallingii, W.F.

    2011-01-01

    Detailed information on plant penetration activities by pear psylla Cacopsylla pyri L. (Hemiptera Psyllidae) is essential to study phytoplasma transmission of “Candidatus Phytoplasma pyri” responsible of pear decline disease (PD) and to trace and evaluate resistant traits in new pear tree selections

  1. ATTRACTION OF MALE SUMMERFORM PEAR PSYLLA TO VOLATILES FROM FEMALE PSYLLA: EFFECTS OF FEMALE AGE, MATING STATUS, AND PRESENCE OF HOST PLANT

    Science.gov (United States)

    Pear psylla, Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae), is a pest of pears throughout North America and western Europe. Previous studies in our laboratory showed that males of the overwintering form (winterform morphotype) were attracted to volatiles from pear shoots infested with post-d...

  2. Attraction of Male Winterform Pear Psylla to Female-produced Volatiles and to Female Extracts and Evidence of Male-Male Repellency

    Science.gov (United States)

    Pear psylla, Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae), is a major pest of commercial pears in North America and Europe. Olfactometer trials have shown that males of both the summerform and winterform morphotype are attracted to female-infested host material. Additional work with the su...

  3. Stylet penetration of Cacopsylla pyri; an electrical penetration graph (EPG) study

    NARCIS (Netherlands)

    Civolani, S.; Leis, M.; Grandi, G.; Garzo, E.; Pasqualini, E.; Musacchi, S.; Chicca, M.; Castaldelli, G.; Rossie, M.; Tjallingii, W.F.

    2011-01-01

    Detailed information on plant penetration activities by pear psylla Cacopsylla pyri L. (Hemiptera Psyllidae) is essential to study phytoplasma transmission of “Candidatus Phytoplasma pyri” responsible of pear decline disease (PD) and to trace and evaluate resistant traits in new pear tree selections

  4. NCBI nr-aa BLAST: CBRC-XTRO-01-2352 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-2352 ref|NP_642468.1| YapH protein [Xanthomonas axonopodis pv. citri s...tr. 306] gb|AAM37004.1| YapH protein [Xanthomonas axonopodis pv. citri str. 306] NP_642468.1 0.30 32% ...

  5. NCBI nr-aa BLAST: CBRC-LAFR-01-1946 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-1946 ref|NP_641456.1| hypothetical protein XAC1119 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM35992.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641456.1 0.50 39% ...

  6. NCBI nr-aa BLAST: CBRC-XTRO-01-2343 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-2343 ref|NP_642141.1| filamentous haemagglutinin [Xanthomonas axonopod...is pv. citri str. 306] gb|AAM36677.1| filamentous haemagglutinin [Xanthomonas axonopodis pv. citri str. 306] NP_642141.1 6e-04 27% ...

  7. NCBI nr-aa BLAST: CBRC-LAFR-01-0106 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0106 ref|NP_643310.1| MFS transporter [Xanthomonas axonopodis pv. citr...i str. 306] gb|AAM37846.1| MFS transporter [Xanthomonas axonopodis pv. citri str. 306] NP_643310.1 0.54 26% ...

  8. NCBI nr-aa BLAST: CBRC-PTRO-23-0017 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PTRO-23-0017 ref|NP_642953.1| phage-related terminase [Xanthomonas axonopodis ...pv. citri str. 306] gb|AAM37489.1| phage-related terminase [Xanthomonas axonopodis pv. citri str. 306] NP_642953.1 5.6 33% ...

  9. NCBI nr-aa BLAST: CBRC-LAFR-01-1513 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-1513 ref|NP_643310.1| MFS transporter [Xanthomonas axonopodis pv. citr...i str. 306] gb|AAM37846.1| MFS transporter [Xanthomonas axonopodis pv. citri str. 306] NP_643310.1 1.1 24% ...

  10. NCBI nr-aa BLAST: CBRC-LAFR-01-1577 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-1577 ref|NP_641723.1| hypothetical protein XAC1388 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM36259.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641723.1 0.077 39% ...

  11. NCBI nr-aa BLAST: CBRC-DNOV-01-2969 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2969 ref|NP_641277.1| hypothetical protein XAC0925 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM35813.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641277.1 3.0 27% ...

  12. NCBI nr-aa BLAST: CBRC-ACAR-01-0078 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0078 ref|NP_644793.1| avirulence protein [Xanthomonas axonopodis pv. c...itri str. 306] gb|AAM39311.1| avirulence protein [Xanthomonas axonopodis pv. citri str. 306] NP_644793.1 2e-04 26% ...

  13. NCBI nr-aa BLAST: CBRC-XTRO-01-0707 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0707 ref|NP_644475.1| acetate permease [Xanthomonas axonopodis pv. cit...ri str. 306] gb|AAM39011.1| solute:Na+ symporter [Xanthomonas axonopodis pv. citri str. 306] NP_644475.1 1e-136 79% ...

  14. NCBI nr-aa BLAST: CBRC-DRER-13-0011 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-13-0011 ref|NP_642674.1| hypothetical protein XAC2357 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM37210.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_642674.1 5e-05 30% ...

  15. NCBI nr-aa BLAST: CBRC-XTRO-01-0645 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0645 ref|NP_642163.1| hypothetical protein XAC1837 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM36699.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_642163.1 9e-42 40% ...

  16. NCBI nr-aa BLAST: CBRC-DYAK-01-0032 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DYAK-01-0032 ref|NP_640378.1| D-3-phosphoglycerate dehydrogenase [Xanthomonas ...axonopodis pv. citri str. 306] gb|AAM34914.1| D-3-phosphoglycerate dehydrogenase [Xanthomonas axonopodis pv. citri str. 306] NP_640378.1 1.1 32% ...

  17. NCBI nr-aa BLAST: CBRC-XTRO-01-0625 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0625 ref|NP_642734.1| integral membrane protein [Xanthomonas axonopodi...s pv. citri str. 306] gb|AAM37270.1| integral membrane protein [Xanthomonas axonopodis pv. citri str. 306] NP_642734.1 1e-62 27% ...

  18. NCBI nr-aa BLAST: CBRC-ACAR-01-0713 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0713 ref|NP_641889.1| glucose-galactose transporter [Xanthomonas axono...podis pv. citri str. 306] gb|AAM36425.1| glucose-galactose transporter [Xanthomonas axonopodis pv. citri str. 306] NP_641889.1 4.7 35% ...

  19. NCBI nr-aa BLAST: CBRC-BTAU-01-1628 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-1628 ref|NP_641751.1| phosphatidate cytidylyltransferase [Xanthomonas ...axonopodis pv. citri str. 306] gb|AAM36287.1| phosphatidate cytidylyltransferase [Xanthomonas axonopodis pv. citri str. 306] NP_641751.1 6.8 24% ...

  20. NCBI nr-aa BLAST: CBRC-DYAK-06-0020 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DYAK-06-0020 ref|NP_642141.1| filamentous haemagglutinin [Xanthomonas axonopod...is pv. citri str. 306] gb|AAM36677.1| filamentous haemagglutinin [Xanthomonas axonopodis pv. citri str. 306] NP_642141.1 0.001 26% ...

  1. NCBI nr-aa BLAST: CBRC-BTAU-01-2358 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-2358 ref|NP_641879.1| hypothetical protein XAC1546 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM36415.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641879.1 5.2 31% ...

  2. NCBI nr-aa BLAST: CBRC-ACAR-01-0078 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0078 ref|NP_644725.1| avirulence protein [Xanthomonas axonopodis pv. c...itri str. 306] gb|AAM39243.1| avirulence protein [Xanthomonas axonopodis pv. citri str. 306] NP_644725.1 2e-04 26% ...

  3. NCBI nr-aa BLAST: CBRC-DRER-13-0009 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-13-0009 ref|NP_642674.1| hypothetical protein XAC2357 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM37210.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_642674.1 5e-05 30% ...

  4. NCBI nr-aa BLAST: CBRC-DDIS-03-0106 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DDIS-03-0106 ref|NP_643514.1| hypothetical protein XAC3206 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM38050.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_643514.1 2e-31 30% ...

  5. NCBI nr-aa BLAST: CBRC-DNOV-01-3199 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-3199 ref|NP_641280.1| extracellular protease [Xanthomonas axonopodis p...v. citri str. 306] gb|AAM35816.1| extracellular protease [Xanthomonas axonopodis pv. citri str. 306] NP_641280.1 5.1 25% ...

  6. NCBI nr-aa BLAST: CBRC-GGAL-35-0330 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GGAL-35-0330 ref|NP_641879.1| hypothetical protein XAC1546 [Xanthomonas axonop...odis pv. citri str. 306] gb|AAM36415.1| conserved hypothetical protein [Xanthomonas axonopodis pv. citri str. 306] NP_641879.1 0.034 26% ...

  7. NCBI nr-aa BLAST: CBRC-ETEL-01-0961 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ETEL-01-0961 ref|NP_644405.1| dipeptidyl peptidase [Xanthomonas axonopodis pv.... citri str. 306] gb|AAM38941.1| dipeptidyl peptidase [Xanthomonas axonopodis pv. citri str. 306] NP_644405.1 0.67 54% ...

  8. Wavelength and polarization affect phototaxis of the Asian citrus psyllid

    Science.gov (United States)

    The Asian citrus psyllid, D. citri, is a primary pest for citrus production due to its status as a vector of the citrus disease, huanglongbing. We investigated phototactic behavior of D. citri to evaluate effects of light of specific wavelength or polarization using a horizontal bioassay arena. Wave...

  9. Identification of gene candidates associated with huanglongbing tolerance using Candidatus Liberibacter asiaticus flagellin 22 as a proxy to challenge citrus

    Science.gov (United States)

    Plant defense elicited by pathogen-associated molecular patterns (PAMPs) is an important component of disease resistance. Previous research indicated the canker resistance in citrus correlates with responsiveness to Xcc-flg22, the 22 amino acid PAMP from the flagellin of Xanthomonas citri ssp. citri...

  10. NCBI nr-aa BLAST: CBRC-DNOV-01-1305 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-1305 ref|NP_640667.1| vanillate O-demethylase oxygenase subunit [Xanth...omonas axonopodis pv. citri str. 306] gb|AAM35203.1| vanillate O-demethylase oxygenase subunit [Xanthomonas axonopodis pv. citri str. 306] NP_640667.1 5.1 38% ...

  11. Bioassay on oviposition repellency of non-preferable plant extracts against citrus red mite Panonychus citri%非嗜食植物提取物对桔全爪螨产卵的驱避性测定

    Institute of Scientific and Technical Information of China (English)

    岑伊静; 庞雄飞; 周琼; 彭跃峰; 徐长宝

    2004-01-01

    The oviposition repellency of the alcohol extracts from 50 species of non-preferable plants and azadirachtin against citrus red mite ( Panonychus cirri ) was determined using laboratory bioassays. In choice tests, the extracts from 42 of the 50 plant species and 1% azadirachtin (2000×) significantly reduced oviposition 1d after treatment.The repellency effect of the extracts from the 42 plant species was better than that of azadirachtin. Mikania micrantha extract had the best result, with an Interference Index of Population Control (IIPC) of 0.087 1 d after treatment, and significantly reduced oviposition for 9 d, while the extracts from Sesbania cannabina, Allium tuberosum , Paederia scandens , Duranta erecta and Dicranopteris pedata also had good effects, with an oviposition repellency of over 70 % 1 d after treatment, and significantly reduced oviposition for 4 - 6 d. The effect became weaker as time went on. None of the extracts showed significant oviposition attraction.

  12. 复方川贝止咳糖浆中化橘红和薄荷脑的薄层色谱鉴别%Identification of Exocarpium Citri Grandis and Mentholum in Compound Bulbus Fritillariae Cough Syrup by TLC

    Institute of Scientific and Technical Information of China (English)

    辛振杰; 李依信; 李永斌

    2007-01-01

    目的 提高复方川贝止咳糖浆的质量标准,更好地控制产品的质量.方法 采用薄层色谱(TLC)法,对制剂中化橘红和薄荷脑的定性方法进行研究.结果 建立了两味药材的TLC鉴别方法,且方法专属性强,操作简便可靠,便于普通检验室操作.结论 TLC法专属性强,可用于复方川贝止咳糖浆的质量控制.

  13. Bioactivity of the Mixture of Two Botanical Pesticide against Panonychus citri%两种植物提取物不同比例混合对柑橘全爪螨的毒杀活性

    Institute of Scientific and Technical Information of China (English)

    刘浩强; 冉春; 李鸿筠; 胡军华; 姚廷山

    2010-01-01

    试验测定了使君子Quisquabis indical和巴豆Croton tiglium在碱水溶液中的提取物对柑橘全爪螨的毒杀活性,并且测定了使君子和巴豆不同比例混合物对柑橘全爪螨的毒杀作用和增效作用.测定结果表明使君子和巴豆的碱水提取液在10 mg/L时单独使用对柑橘全爪螨24 h的校正死亡率分别为94.50%、100%,不同比例混合试验显示:对柑橘全爪螨触杀毒性最强的是使君子和巴豆提取物以1:2的比例混合的混合物.其LC50值和LC95值分别为0.8545、2.9500 mg/L,混剂的供毒系数为270.2589.此外当使君子和巴豆的提取物以1:1的比例混合时,其LC50值和LC95值分别为0.8150、3.7843 mg/L,混剂的供毒系数为279.8057,说明使君子和巴豆以1:1到2:1之间的比例混合时有较强的增效作用.

  14. Sensitivity Analysis of qPCR in Detecting the Xanthomonas axonopodis pv.citri%利用qPCR技术检测柑橘溃疡病菌的灵敏度分析

    Institute of Scientific and Technical Information of China (English)

    李文娟; 肖翠; 戴素明; 李大志; 邓子牛

    2015-01-01

    以常规PCR为对照,分析了实时荧光定量PCR (qPCR)检测柑橘溃疡病菌DNA和柑橘溃疡病菌菌量的灵敏度,并对接种柑橘溃疡病菌后的冰糖橙样品进行了动态监测.结果显示,qPCR检测柑橘溃疡病菌DNA和柑橘溃疡病菌菌量的灵敏度比常规PCR高l~3个数量级,且常规PCR对柑橘溃疡病菌DNA的检测会受到寄主植物DNA的干扰,qPCR不受寄主植物DNA影响;对接种后冰糖橙样品的分析,qPCR即时就能检测到柑橘溃疡病菌,并能实时观察到柑橘溃疡病菌在寄主叶片的增殖情况,而常规PCR要接种后2d才能检测到柑橘溃疡病菌.

  15. 橘光绿天牛危害九里香的现状调查及化学防治试验%Investigation on Chelidonium citri harm to Murraya paniculata and experiment on its chemical control

    Institute of Scientific and Technical Information of China (English)

    吴清荣

    2007-01-01

    本文对橘光绿天牛蛀食九里香枝干造成枝干空洞、枯死的现状进行调查,并应用注射法进行不同药品和浓度的化学防治试验.结果表明,橘光绿天牛对九里香的危害程度达62.5%.化学防治中,除了40%氧化乐果乳油80 g·L-1的处理外,各处理的虫口平均死亡率达80%以上,综合药效、环保、经济考量,选用80%敌敌畏乳油80 g·L-1保绿宁乳油80 g·L-1是防治九里香橘光绿天牛危害最佳选择.

  16. Mycoplasma infections of plants.

    Science.gov (United States)

    Bove, J M

    1981-07-01

    Plants can be infected by two types of wall-less procaryotes, spiroplasmas and mycoplasma-like organisms (MLO), both located intracellularly in the phloem tissues of affected plants. Spiroplasmas have been cultured, characterized and shown to be true members of the class Mollicutes. MLO have not yet been cultured or characterized; they are thought to be mycoplasma-like on the basis of their ultrastructure as seen in situ, their sensitivity to tetracycline and resistance to penicillin. Mycoplasmas can also be found on the surface of plants. These extracellularly located organisms are members of the following genera: Spiroplasma. Mycoplasma and Acholeplasma. The presence of such surface mycoplasmas must not be overlooked when attempts to culture MLO from affected plants are undertaken. Sensitive serological techniques such as the enzyme-linked immunosorbent assay (ELISA) can successfully be used to compare the MLO located in the phloem of affected plants with those eventually cultured from the same plants. In California and Morocco periwinkles naturally infected with both Spiroplasma citri and MLO have been reported. With such doubly infected plants, the symptom expression has been that characteristic of the MLO disease (phyllody or stolbur), not that given by S. citri. Only S. citri can be cultured from such plants, but this does not indicate that S. citri is the causal agent of the disease expressed by the plant. In California many nonrutaceous plants have been found to be infected with S. citri. Stubborn affected citrus trees represent an important reservoir of S. citri, and Circulifer tenellus is an active leafhopper vector of S. citri. Hence, it is not surprising that in California MLO-infected fruit trees could also become infected with S. citri but it would not mean that S. citri is the causal agent of the disease. Criteria are discussed that are helpful in distinguishing between MLO infections and S. citri infections.

  17. Bacterial diversity analysis of Huanglongbing pathogen-infected citrus, using PhyloChip and 16S rRNA gene clone library sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Shankar Sagaram, U.; DeAngelis, K.M.; Trivedi, P.; Andersen, G.L.; Lu, S.-E.; Wang, N.

    2009-03-01

    host plants (5,27,40). Tatineni and colleagues discovered that the HLB bacteria were unevenly distributed in phloem of bark tissue, vascular tissue of the leaf midrib, roots, and different floral and fruit parts (43). Unsuccessful attempts in culturing the pathogen are notably hampering efforts to understand its biology and pathogenesis mechanism. Using a modified Koch's Postulates approach, Jagoueix and colleagues were able to re-infect periwinkle plants from a mixed microbial community harvested from HLB diseased plants (25). Emergence of the disease in otherwise healthy plants led to the conclusion that HLB was associated with Candidatus Liberibacter sp. based on its 16S rDNA sequence (18,25). Currently, three species of the pathogen are recognized from trees with HLB disease based on 16S rDNA sequence: Ca. Liberibacter asiaticus (Las), Ca. Liberibacter africanus (Laf), and Ca. Liberibacter americanus (Lam); Las is the most prevalent species among HLB diseased trees (5,12,18,25,44). Las is naturally transmitted to citrus by the psyllid, Diaphorina citri Kuwayama, and can be artificially transmitted by grafting from citrus to citrus and dodder (Cuscuta campestris) to periwinkle (Catharanthus roseus) or tobacco (Nicotiana tabacum Xanthi) (5). Based on current research regarding the associations of Liberibacter in planta there is not enough evidence to implicate Liberibacter as the definitive causal agent of HLB disease due to its resistance to cultivation in vitro. It is possible that HLB disease may be the result of complex etiology where Liberibacter interacts with other endophytic bacteria. However, there is not enough evidence regarding its association(s) in planta to make this conclusion, nor is it known whether associated microbial communities play a role in expression of pathogenic traits. The main objective of the study was to test the hypothesis that other bacteria besides Ca. Liberibacter spp. are associated with citrus greening disease. The differences

  18. Bacterial diversity analysis of Huanglongbing pathogen-infected citrus, using PhyloChip and 16S rRNA gene clone library sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Shankar Sagaram, U.; DeAngelis, K.M.; Trivedi, P.; Andersen, G.L.; Lu, S.-E.; Wang, N.

    2009-03-01

    host plants (5,27,40). Tatineni and colleagues discovered that the HLB bacteria were unevenly distributed in phloem of bark tissue, vascular tissue of the leaf midrib, roots, and different floral and fruit parts (43). Unsuccessful attempts in culturing the pathogen are notably hampering efforts to understand its biology and pathogenesis mechanism. Using a modified Koch's Postulates approach, Jagoueix and colleagues were able to re-infect periwinkle plants from a mixed microbial community harvested from HLB diseased plants (25). Emergence of the disease in otherwise healthy plants led to the conclusion that HLB was associated with Candidatus Liberibacter sp. based on its 16S rDNA sequence (18,25). Currently, three species of the pathogen are recognized from trees with HLB disease based on 16S rDNA sequence: Ca. Liberibacter asiaticus (Las), Ca. Liberibacter africanus (Laf), and Ca. Liberibacter americanus (Lam); Las is the most prevalent species among HLB diseased trees (5,12,18,25,44). Las is naturally transmitted to citrus by the psyllid, Diaphorina citri Kuwayama, and can be artificially transmitted by grafting from citrus to citrus and dodder (Cuscuta campestris) to periwinkle (Catharanthus roseus) or tobacco (Nicotiana tabacum Xanthi) (5). Based on current research regarding the associations of Liberibacter in planta there is not enough evidence to implicate Liberibacter as the definitive causal agent of HLB disease due to its resistance to cultivation in vitro. It is possible that HLB disease may be the result of complex etiology where Liberibacter interacts with other endophytic bacteria. However, there is not enough evidence regarding its association(s) in planta to make this conclusion, nor is it known whether associated microbial communities play a role in expression of pathogenic traits. The main objective of the study was to test the hypothesis that other bacteria besides Ca. Liberibacter spp. are associated with citrus greening disease. The differences

  19. Characterization of Xanthomonas spp. strains by bacteriocins Caracterização de isolados de Xanthomonas spp. por bacterocinas

    Directory of Open Access Journals (Sweden)

    Marcel Bonini

    2007-03-01

    Full Text Available Twenty-five strains of Xanthomonas axonopodis pv. citri and 14 strains of Xanthomonas spp. were tested for bacteriocin production. X. axonopodis pv. passiflorae strains were sensitive to the bacteriocins produced by the 25 X. axonopodis pv. citri strains evaluated in this study while strains of X. axonopodis pv. manihotis and X. campestris pv. campestris showed variable sensitivity. Only five of the 25 X. axonopodis pv. citri strains were not inhibited by the bacteriocins produced by the two X. axonopodis pv. passiflorae strains. The bacteriocins produced by the Xanthomonas axonopodis pv. citri (FDC-806 and X. axonopodis pv. passiflorae (Mar-2850 A strains were thermolabile, resistant to lysozyme and sensitive to DNAse. The bacteriocin produced by X. axonopodis pv. passiflorae was resistant to the action of proteinase K, trypsin and RNAse while the bacteriocin produced by X. axonopodis pv. citri was sensitive to these enzymes. The bacteriocins produced by X. axonopodis pv. passiflorae and X. axonopodis pv. citri were called passifloricin and citricin, respectively.Vinte e cinco isolados de Xanthomonas axonopodis pv. citri e 14 isolados de Xanthomonas spp. foram comparados a fim de verificar a capacidade de produção de bacteriocina e a sua sensibilidade. Isolados de X. axonopodis pv. passiflorae foram sensíveis às bacteriocinas produzidas por 25 isolados de X. axonopodis pv. citri avaliados e os isolados de X. axonopodis pv. manihotis e X. campestris pv. campestris apresentaram sensibilidade variável. Dos 25 isolados de X. axonopodis pv. citri apenas cinco não foram inibidos pelas bacteriocinas produzidas por dois isolados de X. axonopodis pv. passiflorae. As bacteriocinas produzidas pelos isolados de X. axonopodis pv. citri (FDC-806 e de X. axonopodis pv. passiflorae (Mar-2850 A foram termolábeis e resistentes à lisozima e sensíveis a DNAse. A bacteriocina produzida pelo isolado de X. axonopodis pv. passiflorae foi resistente à a

  20. INTRODUCTION Curing is holding fruits at temperatures and ...

    African Journals Online (AJOL)

    One of the. EFFECT OF CURING ON THE SHELF LIFE OF AMBERSWEET ORANGES ... Alternaria citri. Incidence was lowest in fruits cured in the shade to 7% weight loss. .... Period of Storage (Weeks)/ Incidence of Decay (%). (% Wt. loss). 1.

  1. Use of molecular tools for the identification of males of some scale insects (Hemiptera: Coccoidea, in pheromone traps used for monitoring and comparison with females

    Directory of Open Access Journals (Sweden)

    István Tóbiás

    2010-11-01

    Full Text Available Species from Pseudococcidae family were studied. It was determined that the dry males of Planococcus citri, and Pseudococcus comstocki, collected by pheromone traps could be useful for the molecular analyses too. The ITS-2 sequences of males and females in case of Pl.citri, Planococcus ficus and Ps. comstocki were identical. This molecular method could differentiate the two mealybug species and this method can be useful to have idea specimens collected by pheromone traps.

  2. Antibacterial activity of cyclo(L-Pro-L-Tyr) and cyclo(D-Pro-L-Tyr) from Streptomyces sp. strain 22-4 against phytopathogenic bacteria.

    Science.gov (United States)

    Wattana-Amorn, Pakorn; Charoenwongsa, Waranya; Williams, Christopher; Crump, Matthew P; Apichaisataienchote, Busaya

    2016-09-01

    Two bioactive cyclic dipeptides, cyclo(L-Pro-L-Tyr) and cyclo(D-Pro-L-Tyr), were isolated from the culture broth of Streptomyces sp. strain 22-4 and tested against three economically important plant pathogens, Xanthomonas axonopodis pv. citri, Ralstonia solanacearum and Clavibacter michiganensis. Both cyclic dipeptides were active against X. axonopodis pv. citri and R. Solanacearum with MIC of 31.25 μg/mL. No activity could be observed against C. michiganensis.

  3. Comparative genomic analysis of Xanthomonas axonopodis pv. citrumelo F1, which causes citrus bacterial spot disease, and related strains provides insights into virulence and host specificity.

    Science.gov (United States)

    Jalan, Neha; Aritua, Valente; Kumar, Dibyendu; Yu, Fahong; Jones, Jeffrey B; Graham, James H; Setubal, João C; Wang, Nian

    2011-11-01

    Xanthomonas axonopodis pv. citrumelo is a citrus pathogen causing citrus bacterial spot disease that is geographically restricted within the state of Florida. Illumina, 454 sequencing, and optical mapping were used to obtain a complete genome sequence of X. axonopodis pv. citrumelo strain F1, 4.9 Mb in size. The strain lacks plasmids, in contrast to other citrus Xanthomonas pathogens. Phylogenetic analysis revealed that this pathogen is very close to the tomato bacterial spot pathogen X. campestris pv. vesicatoria 85-10, with a completely different host range. We also compared X. axonopodis pv. citrumelo to the genome of citrus canker pathogen X. axonopodis pv. citri 306. Comparative genomic analysis showed differences in several gene clusters, like those for type III effectors, the type IV secretion system, lipopolysaccharide synthesis, and others. In addition to pthA, effectors such as xopE3, xopAI, and hrpW were absent from X. axonopodis pv. citrumelo while present in X. axonopodis pv. citri. These effectors might be responsible for survival and the low virulence of this pathogen on citrus compared to that of X. axonopodis pv. citri. We also identified unique effectors in X. axonopodis pv. citrumelo that may be related to the different host range as compared to that of X. axonopodis pv. citri. X. axonopodis pv. citrumelo also lacks various genes, such as syrE1, syrE2, and RTX toxin family genes, which were present in X. axonopodis pv. citri. These may be associated with the distinct virulences of X. axonopodis pv. citrumelo and X. axonopodis pv. citri. Comparison of the complete genome sequence of X. axonopodis pv. citrumelo to those of X. axonopodis pv. citri and X. campestris pv. vesicatoria provides valuable insights into the mechanism of bacterial virulence and host specificity.

  4. Comparative Genomic Analysis of Xanthomonas axonopodis pv. citrumelo F1, Which Causes Citrus Bacterial Spot Disease, and Related Strains Provides Insights into Virulence and Host Specificity ▿ #

    Science.gov (United States)

    Jalan, Neha; Aritua, Valente; Kumar, Dibyendu; Yu, Fahong; Jones, Jeffrey B.; Graham, James H.; Setubal, João C.; Wang, Nian

    2011-01-01

    Xanthomonas axonopodis pv. citrumelo is a citrus pathogen causing citrus bacterial spot disease that is geographically restricted within the state of Florida. Illumina, 454 sequencing, and optical mapping were used to obtain a complete genome sequence of X. axonopodis pv. citrumelo strain F1, 4.9 Mb in size. The strain lacks plasmids, in contrast to other citrus Xanthomonas pathogens. Phylogenetic analysis revealed that this pathogen is very close to the tomato bacterial spot pathogen X. campestris pv. vesicatoria 85-10, with a completely different host range. We also compared X. axonopodis pv. citrumelo to the genome of citrus canker pathogen X. axonopodis pv. citri 306. Comparative genomic analysis showed differences in several gene clusters, like those for type III effectors, the type IV secretion system, lipopolysaccharide synthesis, and others. In addition to pthA, effectors such as xopE3, xopAI, and hrpW were absent from X. axonopodis pv. citrumelo while present in X. axonopodis pv. citri. These effectors might be responsible for survival and the low virulence of this pathogen on citrus compared to that of X. axonopodis pv. citri. We also identified unique effectors in X. axonopodis pv. citrumelo that may be related to the different host range as compared to that of X. axonopodis pv. citri. X. axonopodis pv. citrumelo also lacks various genes, such as syrE1, syrE2, and RTX toxin family genes, which were present in X. axonopodis pv. citri. These may be associated with the distinct virulences of X. axonopodis pv. citrumelo and X. axonopodis pv. citri. Comparison of the complete genome sequence of X. axonopodis pv. citrumelo to those of X. axonopodis pv. citri and X. campestris pv. vesicatoria provides valuable insights into the mechanism of bacterial virulence and host specificity. PMID:21908674

  5. Evidence for gene flow between two sympatric mealybug species (Insecta; Coccoidea; Pseudococcidae).

    Science.gov (United States)

    Kol-Maimon, Hofit; Ghanim, Murad; Franco, José Carlos; Mendel, Zvi

    2014-01-01

    Occurrence of inter-species hybrids in natural populations might be evidence of gene flow between species. In the present study we found evidence of gene flow between two sympatric, genetically related scale insect species--the citrus mealybug Planococcus citri (Risso) and the vine mealybug Planococcus ficus (Signoret). These species can be distinguished by morphological, behavioral, and molecular traits. We employed the sex pheromones of the two respective species to study their different patterns of male attraction. We also used nuclear ITS2 (internal transcribed spacer 2) and mitochondrial COI (Cytochrome c oxidase sub unit 1) DNA sequences to characterize populations of the two species, in order to demonstrate the outcome of a possible gene flow between feral populations of the two species. Our results showed attraction to P. ficus pheromones of all tested populations of P. citri males but not vice versa. Furthermore, ITS2 sequences revealed the presence of 'hybrid females' among P. citri populations but not among those of P. ficus. 'hybrid females' from P. citri populations identified as P. citri females according to COI sequences. We offer two hypotheses for these results. 1) The occurrence of phenotypic and genotypic traits of P. ficus in P. citri populations may be attributed to both ancient and contemporary gene flow between their populations; and 2) we cannot rule out that an ancient sympatric speciation by which P. ficus emerged from P. citri might have led to the present situation of shared traits between these species. In light of these findings we also discuss the origin of the studied species and the importance of the pherotype phenomenon as a tool with which to study genetic relationships between congener scale insects.

  6. Evidence for gene flow between two sympatric mealybug species (Insecta; Coccoidea; Pseudococcidae.

    Directory of Open Access Journals (Sweden)

    Hofit Kol-Maimon

    Full Text Available Occurrence of inter-species hybrids in natural populations might be evidence of gene flow between species. In the present study we found evidence of gene flow between two sympatric, genetically related scale insect species--the citrus mealybug Planococcus citri (Risso and the vine mealybug Planococcus ficus (Signoret. These species can be distinguished by morphological, behavioral, and molecular traits. We employed the sex pheromones of the two respective species to study their different patterns of male attraction. We also used nuclear ITS2 (internal transcribed spacer 2 and mitochondrial COI (Cytochrome c oxidase sub unit 1 DNA sequences to characterize populations of the two species, in order to demonstrate the outcome of a possible gene flow between feral populations of the two species. Our results showed attraction to P. ficus pheromones of all tested populations of P. citri males but not vice versa. Furthermore, ITS2 sequences revealed the presence of 'hybrid females' among P. citri populations but not among those of P. ficus. 'hybrid females' from P. citri populations identified as P. citri females according to COI sequences. We offer two hypotheses for these results. 1 The occurrence of phenotypic and genotypic traits of P. ficus in P. citri populations may be attributed to both ancient and contemporary gene flow between their populations; and 2 we cannot rule out that an ancient sympatric speciation by which P. ficus emerged from P. citri might have led to the present situation of shared traits between these species. In light of these findings we also discuss the origin of the studied species and the importance of the pherotype phenomenon as a tool with which to study genetic relationships between congener scale insects.

  7. 三种季酮酸类对柑桔红蜘蛛室内毒力测定及田间药效试验%Toxicity and field control efficacy of three tetronic acid to citrus red mite Panonychus citri (McGregor) (Arachnida: Tetranychidae)

    Institute of Scientific and Technical Information of China (English)

    邓福新

    2013-01-01

    测试3种季酮酸类药剂对脐橙上柑桔红蜘蛛的室内毒力和田间药效.结果表明:柑桔红蜘蛛对螺螨酯、螺甲螨酯和螺虫乙酯三种药剂对的敏感性远低于阿维菌素,以田间喷施不同药剂2 d后以阿维菌素的防效最高,螺虫乙酯为最低且差异显著.7d后螺螨酯、螺甲螨酯和阿维菌素的防效无显著差异,但这三种药的防效仍然显著高于螺虫乙酯.而到14 d后,四种药剂的防效相当,无显著差异.

  8. Toxicity of L1-2 fraction of Arctium lappa against Panonychus citri (McGregor) (Acari :Tetranychidae) and its effects on several metabolic enzymes%牛蒡L1-2组分对桔全爪螨的毒性和几种代谢酶的作用

    Institute of Scientific and Technical Information of China (English)

    胡军华; 马丽娜; 冉春; 李鸿筠; 姚廷山; 刘浩强; 雷慧德

    2010-01-01

    [目的]探讨杀螨植物牛蒡Arctium lappa L.提取物中主要杀螨成分L1-2的杀螨作用机理.[方法]采用叶片浸渍法处理桔全爪螨Panonychus citri雌成螨后,测定了静止期、兴奋期、痉挛期、麻痹期、复苏和死亡期5个中毒阶段试虫体内几种代谢酶的活性.[结果]L1-2组分在静止期和复苏期对羧酸酯酶(carboxylesterase,CarE)具有一定的抑制作用,在其他时期均激活CarE活性.除了静止期外,在其他时期均能激活乙酰胆碱酯酶(acetylcholinesterase,AChE)和谷胱甘肽转移酶(glutathione S-transferases,GSTs)的活性,在痉挛期和麻痹期活性增强,随后在麻痹期和复苏期降低.[结论]L1-2组分对CarE的抑制与其毒杀活性有关,而中毒试虫的复苏可能与AChE和GSTs有关.该组分可在较长时期内影响桔全爪螨的神经传导及消化和生殖系统,具有潜在的应用研究价值.

  9. [First record of Platycorypha nigrivirga Burckhardt (Hemiptera: Psylloidea), in Tipuana tipu (Benth.), from Brazil].

    Science.gov (United States)

    Santana, Dalva L Q; Burckhardt, Daniel; Aguiar, António M F

    2006-01-01

    The occurrence of Platycorypha nigrivirga Burckhardt (Hemiptera: Psyllidae, Acizzinae), is reported for the first time, in Brazil. The psyllid was observed causing injuries on leaves and shoots of Tipuana tipu (Benth.), in urban areas of Paraná State. P. nigrivirga are tiny sap-sucking insects that feed on phloem and cause curling, stains and fall of leaves. The excrements of these insects accumulate on the leaves propitiating fungus growth on the leaves, or they fall in great amount on the cars dirtying them as if they were sparkling of white and sticky ink.

  10. Adaptation of genetically monomorphic bacteria: evolution of copper resistance through multiple horizontal gene transfers of complex and versatile mobile genetic elements.

    Science.gov (United States)

    Richard, D; Ravigné, V; Rieux, A; Facon, B; Boyer, C; Boyer, K; Grygiel, P; Javegny, S; Terville, M; Canteros, B I; Robène, I; Vernière, C; Chabirand, A; Pruvost, O; Lefeuvre, P

    2017-04-01

    Copper-based antimicrobial compounds are widely used to control plant bacterial pathogens. Pathogens have adapted in response to this selective pressure. Xanthomonas citri pv. citri, a major citrus pathogen causing Asiatic citrus canker, was first reported to carry plasmid-encoded copper resistance in Argentina. This phenotype was conferred by the copLAB gene system. The emergence of resistant strains has since been reported in Réunion and Martinique. Using microsatellite-based genotyping and copLAB PCR, we demonstrated that the genetic structure of the copper-resistant strains from these three regions was made up of two distant clusters and varied for the detection of copLAB amplicons. In order to investigate this pattern more closely, we sequenced six copper-resistant X. citri pv. citri strains from Argentina, Martinique and Réunion, together with reference copper-resistant Xanthomonas and Stenotrophomonas strains using long-read sequencing technology. Genes involved in copper resistance were found to be strain dependent with the novel identification in X. citri pv. citri of copABCD and a cus heavy metal efflux resistance-nodulation-division system. The genes providing the adaptive trait were part of a mobile genetic element similar to Tn3-like transposons and included in a conjugative plasmid. This indicates the system's great versatility. The mining of all available bacterial genomes suggested that, within the bacterial community, the spread of copper resistance associated with mobile elements and their plasmid environments was primarily restricted to the Xanthomonadaceae family. © 2017 John Wiley & Sons Ltd.

  11. Scientific Opinion on a technical file submitted by the Japanese Authorities to support a derogation request from the EU import requirements for bonsai and topiary trees that are host plants of Anoplophora chinensis

    DEFF Research Database (Denmark)

    Baker, R.; Candresse, T.; Dormannsné Simon, E.

    2010-01-01

    Following a request from the EU Commission, the EFSA PLH Panel conducted a scientific opinion on risk analysis and supporting documents provided by APHIS/USDA in support of the request to remove the Union's plant health import requirement that citrus fruit imported into the EU be sourced from...... groves where, since the beginning of the last cycle of vegetation no symptoms of citrus canker were observed, neither in their vicinities. The PHL Panel concluded that the transmission of Xanthomonas citri subsp. citri (Xcc) on asymptomatic citrus fruit was more likely when the fruit were collected from...

  12. Scientific Opinion on the request from the USA regarding export of Florida citrus fruit to the EU

    DEFF Research Database (Denmark)

    Baker, R.; Candresse, T.; Dormannsné Simon, E.

    2011-01-01

    Following a request from the EU Commission, the EFSA PLH Panel conducted a scientific opinion on risk analysis and supporting documents provided by APHIS/USDA in support of the request to remove the Union's plant health import requirement that citrus fruit imported into the EU be sourced from...... groves where, since the beginning of the last cycle of vegetation no symptoms of citrus canker were observed, neither in their vicinities. The PHL Panel concluded that the transmission of Xanthomonas citri subsp. citri (Xcc) on asymptomatic citrus fruit was more likely when the fruit were collected from...

  13. The effect of nutritional spray programs applied to mitigate symptoms of Huanglongbing on fruit drop caused by HLB and citrus canker on ‘Hamlin’ orange trees

    OpenAIRE

    Roberts, P. D.; Rouse, R. E.; Teems, S. S.; Sytsma, R. E.; Shobert, Z.

    2014-01-01

    Huanglongbing (HLB) was detected in Florida in 2005 and has reached 100% incidence in certain citrus plantings in southwest Florida. The putative causal agent of HLB in Florida is the bacterium Candidatus Liberibacter asiaticus (CLa).  Citrus canker caused by the bacterium Xanthomonas citri subsp. citri is endemic in Florida.  In 2011 and 2012, fruit drop on young ‘Hamlin’ trees with symptoms of HLB and/or citrus canker was particularly severe, with more than 90% fruit drop recorded. Nutritio...

  14. Dicty_cDB: CHF836 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 33 7.6 AL844505_76( AL844505 |pid:none) Plasmodium falciparum 3D7 chromos... 33 7.6 DQ167129_1( DQ167129 |pid:none) Infectious bronc...hitis virus isolat... 33 7.6 AM285320_11( AM285320 |pid:none) Spiroplasma citri GII

  15. Survey of current crop management practices in a mixed-ricefield landscape, Mekong Delta, Vietnam - potential of habitat manipulation for improved control of citrus leafminer and citrus red mite

    NARCIS (Netherlands)

    Mele, van P.; Lenteren, van J.C.

    2002-01-01

    In the Mekong Delta, Vietnam, the citrus leafminer Phyllocnistis citrella (CLM) and the citrus red mite Panonychus citri are major pests in both sweet orange (Citrus sinensis) and Tieu mandarin (C. reticulata). Survey data indicate that these pest problems might be aggravated after farmers have comp

  16. Xcc-facilitated agroinfiltration of citrus leaves: a tool for rapid functional analysis of transgenes in citrus leaves.

    Science.gov (United States)

    Jia, Hongge; Wang, Nian

    2014-12-01

    Xanthomonas citri subsp. citri pretreatment before agroinfiltration could significantly promote transient expression in citrus leaves which were previously recalcitrant to agroinfiltration. Transient expression via agroinfiltration is widely used in biotechnology but remains problematic in many economically important plants. Xanthomonas citri subsp. citri (Xcc)-facilitated agroinfiltration was employed to promote transient protein expression in Valencia sweet orange leaves, which are recalcitrant to agroinfiltration. However, it is unclear whether Xcc-facilitated agroinfiltration has broad application, i.e., whether Xcc-facilitated agroinfiltration could be used on other citrus varieties. In addition, we intended to investigate whether Xcc-facilitated agroinfiltration could be used to hasten transgene function assays, e.g., Cre/lox system and Cas9/sgRNA system. In this report, Xcc-facilitated agroinfiltration was further exploited to enhance β-glucuronidase (GUS) expression in five citrus varieties. Xcc-facilitated agroinfiltration also significantly increased GFP expression in six citrus varieties tested. Both GUS and GFP assays indicated that Xcc-facilitated agroinfiltration had the best performance in grapefruit. After Xcc-facilitated agroinfiltration was carried out in grapefruit, protoplast analysis of the transformed cells indicated that there were more than 20 % leaf cells expressing GFP. In grapefruit, usefulness of Xcc-facilitated agroinfiltration was assayed in three case studies: (1) fast functional analysis of Cre/lox system, (2) the heat shock regulation of HSP70B promoter derived from Arabidopsis, and (3) Cas9/sgRNA-mediated genome modification.

  17. Closed-Circuit TV Surveillance Evaluation

    Science.gov (United States)

    2010-12-01

    Canada – Centre for Security Science] Approved by Dr. Paul Chouinard CSS OR Team Leader Approved for release by Dr. Mark Williamson Chair - CSS Document...Interest of Society (CITRIS) and the Samuelson Clinic at the University of California, Berkeley, studied the effectiveness of San Francisco’s small video

  18. The economic importance of acaricides in the control of phytophagous mites and an update on recent acaricide mode of action research

    NARCIS (Netherlands)

    Van Leeuwen, T.; Tirry, L.; Yamamoto, A.; Nauen, R.; Dermauw, W.

    2015-01-01

    Acaricides are one of the cornerstones of an efficient control program for phytophagous mites. An analysis of the global acaricide market reveals that spider mites such as Tetranychus urticae, Panonychus citri and Panonychus ulmi are by far the most economically important species, representing more

  19. Overexpression of a modified plant thionin enhances disease resistance to citrus canker and huanglongbing (HLB, citrus greening)

    Science.gov (United States)

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the United States citrus industry. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized e...

  20. Biological control of mealybugs with lacewing larvae is affected by the presence and type of supplemental prey

    NARCIS (Netherlands)

    Messelink, Gerben J.; Vijverberg, Roland; Leman, Ada; Janssen, Arne

    2016-01-01

    The diversity of prey and food sources in crops has a major effect on biological pest control by generalist predators. In this study, we tested if and how supplemental prey or food affects the control of the citrus mealybug Planococcus citri (Risso) by larvae of the green lacewing Chrysoperla luc